Die rationale Entwicklung von CD5-gerichteten biepitopen CARs mit vollständig humanen Nur-Schwerketten-Antigenerkennungsdomänen
T-Zell-Malignome sind eine Gruppe hämatologischer Krebsarten mit hohen Rezidiv- und Mortalitätsraten. CD5 wird in etwa 85 % der malignen T-Zell-Erkrankungen stark exprimiert, obwohl die normale Expression von CD5 auf Thymozyten, T-Zellen und B1-Zellen beschränkt ist. Die CD5-Expression auf chimären Antigenrezeptor (CAR)-T-Zellen führt jedoch zu CAR-T-Zell-Brudermord. Sobald diese Einschränkung überwunden ist, könnte die auf CD5 gerichtete CAR-T-Therapie eine attraktive Strategie zur Behandlung von malignen T-Zell-Erkrankungen sein.
Hier berichten wir über die Auswahl neuartiger CD5-gerichteter vollständig humaner Schwerketten-Variablendomänen (FHVH) für die Entwicklung eines biepitopen CAR mit der Bezeichnung FHVH3/VH1, das FHVH1 und FHVH3 enthält und für die validiert wurde, dass sie an verschiedene Epitope des CD5-Antigens binden . Um Brudermord in CD5-CAR-T-Zellen zu verhindern, haben wir die Herstellungsverfahren eines CRISPR-Cas9-basierten CD5-Knockouts (CD5KO) und die lentivirale Transduktion von Anti-CD5-CAR optimiert. Funktionelle In-vitro- und In-vivo-Vergleiche zeigten, dass biepitope CD5KO-FHVH3/VH1-CAR-T-Zellen eine verbesserte und länger anhaltende Wirksamkeit zeigten; erzeugte mäßige Zytokin-Sekretion; zeigte ähnliche Spezifitätsprofile wie entweder FHVH1, FHVH3 oder das klinisch getestete H65; und eignet sich daher zur Weiterentwicklung.
Einführung
In den letzten Jahren wurden Fortschritte in der Therapie mit chimären Antigenrezeptor-T-Zellen (CAR-T) erzielt, um maligne B-Zell-Erkrankungen anzugreifen, eine Remission zu induzieren und das langfristige rezidivfreie Überleben bei Patienten mit B-Zell-Leukämie und Lymphom zu verbessern.1, 2 , 3, 4, 5 Die Gesamtprognose von refraktären oder rezidivierenden (r/r) T-Zell-Malignomen ist jedoch im Vergleich zu B-Zell-Malignomen viel schlechter, und Salvage-Chemotherapieschemata bleiben die beste Behandlung für Patienten mit r/r T-Zell-Leukämie oder Lymphom.6, 7, 8 Daher ist es unerlässlich, neuartige wirksame CAR-T-Zelltherapien zur Bekämpfung von malignen T-Zell-Erkrankungen zu entwickeln.
- CD5 wird konstitutiv auf normalen T-Zellen exprimiert und ist in etwa 85 % der malignen T-Zell-Erkrankungen vorhanden.9 Es enthält drei Cystein-reiche Scavenger-Rezeptor-Domänen (SRCR),10, 11, 12 die als inhibitorische Regulatoren sowohl des T-Zell-Rezeptors (TCR ) und B-Zell-Rezeptor (BCR)-Signalisierung und dienen als Ziel für evolutionäre immuntherapeutische Strategien gegen maligne T-Zell-Erkrankungen.13, 14, 15, 16, 17, 18, 19 Darüber hinaus wird CD5 auch häufig in einigen bösartigen wobei die Expression in normalen Geweben auf Thymozyten, T-Zellen und eine kleine Subpopulation von B-Zellen (B1-Zellen) beschränkt ist.20,21
- Die Daten aus einer klinischen Phase-I-Studie , die vom Baylor College of Medicine durchgeführt wurde, haben dies gezeigt CAR-T-Zellen, die ein von der Maus stammendes einzelkettiges variables Fragment (scFv) H65 enthielten, das auf CD5 abzielt, waren sicher und hatten eine Anti-Tumor-Aktivität bei Patienten mit r/r-T-Zell-Malignomen.9 Die Patienten, die eine objektive Reaktion nach Infusion ohne Erhalt geplant hämatopoetische Stammzelltransplantation mit ihrer latenten CD5+-Malignität nach einigen Wochen einen Rückfall erlitten.9 Verbesserungen der Wirksamkeit und Persistenz von CD5-gerichteten CAR-T-Zelltherapien für maligne T-Zell-Erkrankungen sind eindeutig dringend erforderlich.
- Die begrenzte Lebensdauer von CAR-T-Zellen bei Patienten kann mit dem Brudermord von Anti-CD5-CAR-T-Zellen und humanen Anti-Maus-Antikörperantworten zusammenhängen, wohingegen die Anwendung von CD5-Knockout (CD5KO) vollständig humanem (FH) Antikörper-abgeleitetem CAR -T-Zellen können das Überleben und die optimale Funktion von CAR-T-Zellen verlängern.22, 23, 24, 25
- Hier berichten wir über die Auswahl neuer variabler (VH) Domänen der schweren Kette von FH und die rationale Entwicklung einer Fratrizid-resistenten, CD5-gerichteten biepitopen CAR-T-Therapie. Unter Verwendung einer hochwertigen FH-Phagen-Display-Bibliothek, die intern entwickelt wurde und 8,32 × 1010 VH-Domänen enthält, erhielten wir durch Panning, primäres Screening und Identifizierung einige VHs, die spezifisch an rekombinantes CD5-Protein und Zelloberflächen-CD5 binden.
- Wir stellten die Hypothese auf, dass die gleichzeitige Verwendung von VH-Domänen, die auf verschiedene Epitope abzielen, möglicherweise die Funktion von CAR-T-Zellen verbessern und das Risiko eines Tumorausbruchs aufgrund einer Antigenmutation minimieren könnte. Daher wurden durch kompetitive Bindungs-Fluoreszenz-aktivierte Zellsortierung (FACS)-Analyse FHVH1 und FHVH3, zwei VH-Domänen, die an unterschiedliche Epitope von CD5 binden, identifiziert.
- Als nächstes haben wir zur Verlängerung der CAR-T-Zellpersistenz eine 4-1BB co-stimulatorische Domäne im Design des CAR-Konstrukts angewendet, von der berichtet wurde, dass sie die Fähigkeit besitzt, das Überleben von CAR-T-Zellen zu fördern26,27 und anschließend den CD5-Knockout optimiert und lentivirale Transduktionsschritte zur erfolgreichen Generierung von Anti-CD5-CAR-T-Zellen, die Brudermord widerstehen. Darüber hinaus wurden die Funktionen der Degranulation, Zytotoxizität, Zytokinfreisetzung, Expansion und Persistenz von H65-, FHVH1-, FHVH3- und biepitopen FHVH3/VH1-CAR-T-Zellen in vitro und in vivo bestätigt.
- FHVH3/VH1 CAR-T-Zellen zeigten starke und länger anhaltende Wirksamkeits- und Spezifitätsprofile, die denen von klinisch verwendetem H65 ähneln. Dieses biepitope CAR wird derzeit für die weitere Evaluierung und Entwicklung vorbereitet.
Ergebnisse
Screening und Selektion von FH-CD5-spezifischen VH-Domänen
Ein Diagramm des Screening- und Auswahlverfahrens ist in Abbildung 1Adargestellt. Die VH-Domänen wurden aus einer Phagen-Display-Bibliothek, die sowohl naive als auch synthetische menschliche VH-Domänen umfasste, durch drei Protein-Panning-Runden nach der Anreicherung gescreent (Tabelle S1). Insgesamt 92 monoklonale Antikörper wurden für den Enzyme-linked Immunosorbent Assay (ELISA) ausgewählt und unter Verwendung von CD5+ Jurkat- und CD5− Raji-Zellen für das FACS-Screening validiert, von denen 38 Klone spezifisch an das CD5-Protein banden, ohne an andere Kontrollproteine zu binden, und die repräsentative Klone sind in Abbildung 1B gezeigt. Nach der Sequenzierung wurden 29 einzigartige Klone identifiziert. Die Bindungsspezifität dieser 29 Klone mit zusätzlichen CD5+/–-Zelllinien wurde weiter durch FACS-Analyse charakterisiert.
Anti-human CD5 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00480-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD5 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00480-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 325 EUR |
Anti-human CD5 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00480-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-human CD5 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00480-Biotin | BosterBio | 100µg | 234 EUR |
Anti-human CD5 Monoclonal Antibody PerCP-Cy5.5 Conjugated, Flow Validated |
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FC00480-PerCP-Cy5.5 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 360 EUR |
Anti-human CD4 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00344-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD2 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00570-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD7 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC01974-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-Human CD3 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC02675-PE-1 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-Human CD3 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC04405-4-PE | BosterBio | 25 tests, 100 tests, 500 tests | 136.8 EUR |
Anti-Human CD44 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00052-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-Human CD28 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00065-PE | BosterBio | 25 tests, 100 tests | 171.6 EUR |
Anti-Human CD40 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00068-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD14 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00137-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD19 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00154-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD38 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00193-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-Human CD86 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00220-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD59 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00914-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-Human CD81 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC01281-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD55/Daf Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00910-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD8 Cd8a Monoclonal Antibody PE Conjugated, Flow Validated |
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FC02236-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD3 Cd3e Monoclonal Antibody PE Conjugated, Flow Validated |
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FC02675-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD10 MME Monoclonal Antibody PE Conjugated, Flow Validated |
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FC04065-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD117 KIT Monoclonal Antibody PE Conjugated, Flow Validated |
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FC01335-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Mouse Anti-human CD7 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC01974-1-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-Human CD33 Monoclonal Antibody PE-Cy7 Conjugated, Flow Validated |
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FC01508-PE-Cy7 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 363 EUR |
Anti-Human CD54 Icam1 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00171-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD25 IL2RA Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00214-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD45 PTPRC Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00555-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD71 TFRC Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00591-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD62L SELL Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00652-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD90 THY1 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC01818-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 319 EUR |
Anti-human CD13 ANPEP Monoclonal Antibody PE Conjugated, Flow Validated |
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FC02591-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD20 MS4A1 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC03780-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD235a GYPA Monoclonal Antibody PE Conjugated, Flow Validated |
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FC02184-PE | BosterBio | 25µg, 100µg | 395 EUR |
Anti-human CD11b Itgam Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00144-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-Human CD11c ITGAX Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00357-PE | BosterBio | 25 Tests | 290 EUR |
Anti-human CD16 FCGR3A Monoclonal Antibody PE Conjugated, Flow Validated |
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FC01408-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD32 FCGR2A Monoclonal Antibody PE Conjugated, Flow Validated |
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FC01450-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD57 B3GAT1 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC09548-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD33/Siglec 3 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC01508-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-Human HLA-ABC Monoclonal Antibody PE Conjugated, Flow Validated |
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FC20056-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD14 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00137-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 312 EUR |
Anti-human CD19 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00154-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 303 EUR |
Anti-human CD38 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00193-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 303 EUR |
Anti-Human CD86 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00220-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 325 EUR |
Anti-human CD59 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00914-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 325 EUR |
Anti-Human CD81 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC01281-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 325 EUR |
Anti-Human CD3 CD3E Monoclonal Antibody PE-Cy7 Conjugated, Flow Validated |
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FC02675-PE-Cy7 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 341 EUR |
Anti-human CD10 MME Monoclonal Antibody PE-Cy7 Conjugated, Flow Validated |
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FC04065-PE-Cy7 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 338 EUR |
Anti-human CD4 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00344-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 312 EUR |
Anti-human CD4 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00344-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 227 EUR |
Anti-human CD2 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00570-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 312 EUR |
Anti-human CD2 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00570-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-human CD7 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC01974-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 303 EUR |
Anti-human CD7 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC01974-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-Human CD3 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC02675-APC-1 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 312 EUR |
Anti-Human CD3 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC02675-FITC-1 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 227 EUR |
Anti-Human CD3 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC04405-4-APC | BosterBio | 25 tests, 100 tests, 500 tests | 261.6 EUR |
Anti-Human CD3 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC04405-4-FITC | BosterBio | 25 tests, 100 tests, 500 tests | 115.2 EUR |
Anti-Human CD3 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC04405-FITC | BosterBio | 25 tests, 100 tests | 115.2 EUR |
Anti-Human/Mouse CD45R (B220) Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00555-2-PE | BosterBio | 25ug, 100ug | 115.2 EUR |
Anti-Human CD161 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC03917-FITC | BosterBio | 25 tests, 100 tests | 192 EUR |
Anti-Human CD44 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00052-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-Human CD28 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00065-FITC | BosterBio | 25 tests, 100 tests | 157.2 EUR |
Anti-human CD14 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00137-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 208 EUR |
Anti-Human CD14 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00137-FITC-1 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 208 EUR |
Anti-human CD14 Monoclonal Antibody PerCP Conjugated, Flow Validated |
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FC00137-PerCP | BosterBio | 100 Tests, 200 Tests | 334 EUR |
Anti-human CD19 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00154-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 227 EUR |
Anti-human CD19 Monoclonal Antibody PerCP Conjugated, Flow Validated |
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FC00154-PerCP | BosterBio | 25 Tests, 100 Tests, 200 Tests | 338 EUR |
Anti-human CD38 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00193-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 208 EUR |
Anti-Human CD38 Monoclonal Antibody PerCP Conjugated, Flow Validated |
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FC00193-PerCP | BosterBio | 25tests, 100tests, 200tests | 121.2 EUR |
Anti-human CD59 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00914-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-Human CD81 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC01281-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-Human CD62L Sell Monoclonal Antibody PE-Cy7 Conjugated, Flow Validated |
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FC00652-PE-Cy7 | BosterBio | 25tests, 100tests, 200tests | 338 EUR |
Anti-Mouse CD4 Monoclonal Antibody PE-Cyanine7 Conjugated, Flow Validated |
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FC00344-2-PE-Cy7 | BosterBio | 25ug, 100ug | 157.2 EUR |
Anti-Mouse CD4 Monoclonal Antibody PE-Cyanine7 Conjugated, Flow Validated |
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FC00344-PE-Cy7 | BosterBio | 25ug, 100ug | 164.4 EUR |
Anti-human CD4 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00344-Biotin | BosterBio | 100µg | 226 EUR |
Anti-human CD2 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00570-Biotin | BosterBio | 100µg | 234 EUR |
Anti-human CD7 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC01974-Biotin | BosterBio | 100µg | 234 EUR |
Anti-human CD3 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC02675-Biotin | BosterBio | 100µg | 182.4 EUR |
Anti-human CD14 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00137-Biotin | BosterBio | 100µg | 279.6 EUR |
Anti-human CD19 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00154-Biotin | BosterBio | 100µg | 226 EUR |
Anti-human CD38 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00193-Biotin | BosterBio | 100µg | 215 EUR |
Anti-human CD59 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00914-Biotin | BosterBio | 100µg | 264 EUR |
Anti-human CD11b Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00144-Biotin | BosterBio | 100µg | 182.4 EUR |
Anti-Human HLA-DR CD74 Monoclonal Antibody PE-Cy7 Conjugated, Flow Validated |
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FC01340-PE-Cy7 | BosterBio | 25tests, 100tests, 200tests | 338 EUR |
Anti-Human CD14 Monoclonal Antibody mFluor450 Conjugated, Flow Validated |
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FC00137-mFluor450 | BosterBio | 25tests, 100tests, 200tests | 303 EUR |
Anti-Human CD14 Monoclonal Antibody mFluor540 Conjugated, Flow Validated |
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FC00137-mFluor540 | BosterBio | 25tests, 100tests, 200tests | 303 EUR |
Anti-Human CD4 Monoclonal Antibody iFluor647 Conjugated, Flow Validated |
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FC00344-iFluor647 | BosterBio | 25 tests, 100 tests, 200 tests | 303 EUR |
Anti-Human CD4 Monoclonal Antibody mFluor450 Conjugated, Flow Validated |
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FC00344-mFluor450 | BosterBio | 25 tests, 100 tests, 200 tests | 303 EUR |
Anti-Human CD2 Monoclonal Antibody mFluor450 Conjugated, Flow Validated |
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FC00570-mFluor450 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 303 EUR |
Anti-Mouse TER-119 Monoclonal Antibody PE Conjugated, Flow Validated |
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FC30403-PE | BosterBio | 25ug, 100ug | 144 EUR |
Anti-Mouse CD4 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00344-3-FITC | BosterBio | 25ug, 100ug, 500ug | 102 EUR |
Anti-human CD117 KIT Monoclonal Antibody APC Conjugated, Flow Validated |
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FC01335-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 341 EUR |
Mouse anti-human CD7 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC01974-1-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 303 EUR |
Anti-human CD71 TFRC Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00591-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 303 EUR |
Anti-human CD55/Daf Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00910-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 325 EUR |
Anti-human CD55/Daf Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00910-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-human CD90 THY1 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC01818-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 341 EUR |
Anti-human CD8 Cd8a Monoclonal Antibody APC Conjugated, Flow Validated |
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FC02236-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 312 EUR |
Anti-human CD8 Cd8a Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC02236-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 208 EUR |
Anti-Human CD3 Cd3e Monoclonal Antibody APC Conjugated, Flow Validated |
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FC02675-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 312 EUR |
Anti-Human CD3 Cd3e Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC02675-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 227 EUR |
Anti-human CD10 MME Monoclonal Antibody APC Conjugated, Flow Validated |
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FC04065-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 303 EUR |
Anti-human CD10 MME Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC04065-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-Mouse CD19 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00154-1-FITC | BosterBio | 25ug, 100ug, 500ug | 115.2 EUR |
Anti-Mouse CD45.2 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00555-8-FITC | BosterBio | 25ug, 100ug, 500ug | 115.2 EUR |
Anti-Human Fli1 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00399-APC | BosterBio | 100ug | 368 EUR |
Anti-Human VEGI Monoclonal Antibody APC Conjugated, Flow Validated |
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FC02402-APC | BosterBio | Various Sizes | 510 EUR |
Anti-human CD4 Monoclonal Antibody PerCP-Cy5.5 Conjugated, Flow Validated |
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FC00344-PerCP-Cy5.5 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 360 EUR |
Anti-human CD2 Monoclonal Antibody PerCP-Cy5.5 Conjugated, Flow Validated |
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FC00570-PerCP-Cy5.5 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 360 EUR |
Anti-human CD117 KIT Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC01335-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Mouse anti-human CD7 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC01974-1-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-human CD235a GYPA Monoclonal Antibody APC Conjugated, Flow Validated |
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FC02184-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 325 EUR |
Anti-human CD11b Itgam Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00144-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 303 EUR |
Anti-Human CD54 Icam1 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00171-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 325 EUR |
Anti-Human CD54 Icam1 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00171-FITC-1 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-human CD25 IL2RA Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00214-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 312 EUR |
Anti-human CD25 IL2RA Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00214-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 208 EUR |
Anti-human CD45 PTPRC Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00555-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 303 EUR |
Anti-human CD45 PTPRC Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00555-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 208 EUR |
Anti-human CD71 TFRC Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00591-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-human CD62L SELL Monoclonal Antibody APC Conjugated, Flow Validated |
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FC00652-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 325 EUR |
Anti-human CD62L SELL Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00652-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 231 EUR |
Anti-human CD55/Daf Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00910-Biotin | BosterBio | 100µg | 234 EUR |
Anti-human CD32 FCGR2A Monoclonal Antibody APC Conjugated, Flow Validated |
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FC01450-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 338 EUR |
Anti-human CD90 THY1 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC01818-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 268 EUR |
Anti-human CD8 Cd8a Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC02236-Biotin | BosterBio | 100µg | 226 EUR |
Anti-human CD8 Cd8a Monoclonal Antibody PerCP Conjugated, Flow Validated |
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FC02236-PerCP | BosterBio | 25 Tests, 100 Tests, 200 Tests | 338 EUR |
Anti-human CD13 ANPEP Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC02591-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-human CD3 Cd3e Monoclonal Antibody PerCP Conjugated, Flow Validated |
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FC02675-PerCP | BosterBio | 25 Tests, 100 Tests, 200 Tests | 334 EUR |
Anti-human CD20 MS4A1 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC03780-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 303 EUR |
Anti-human CD20 MS4A1 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC03780-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 208 EUR |
Anti-human CD10 MME Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC04065-Biotin | BosterBio | 100µg | 223 EUR |
Anti-human CD15 FUT4 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC04535-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-human CD57 B3GAT1 Monoclonal Antibody APC Conjugated, Flow Validated |
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FC09548-APC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 325 EUR |
Anti-human CD14 Monoclonal Antibody PerCP-Cy5.5 Conjugated, Flow Validated |
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FC00137-PerCP-Cy5.5 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 360 EUR |
Anti-human CD19 Monoclonal Antibody PerCP-Cy5.5 Conjugated, Flow Validated |
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FC00154-PerCP-Cy5.5 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 360 EUR |
Anti-human CD22 Monoclonal Antibody PerCP-Cy5.5 Conjugated, Flow Validated |
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FC01572-PerCP-Cy5.5 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 360 EUR |
Anti-human CD117 KIT Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC01335-Biotin | BosterBio | 100μg | 235 EUR |
Mouse anti-human CD7 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC01974-1-Biotin | BosterBio | 100µg | 234 EUR |
Anti-human CD235a GYPA Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC02184-FITC | BosterBio | 25ug, 100ug | 335 EUR |
Anti-Mouse CD3e Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC04405-3-FITC | BosterBio | 25ug, 100ug, 500ug | 94.8 EUR |
Anti-human HLA-DR HLA-DRA Monoclonal Antibody PE Conjugated, Flow Validated |
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FC00568-PE | BosterBio | 25 Tests, 100 Tests, 200 Tests | 290 EUR |
Anti-human CD11b Itgam Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00144-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 227 EUR |
Anti-human CD11b Itgam Monoclonal Antibody PerCP Conjugated, Flow Validated |
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FC00144-PerCP | BosterBio | 25 Tests, 100 Tests, 200 Tests | 338 EUR |
Anti-human CD25 IL2RA Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00214-Biotin | BosterBio | 100µg | 234 EUR |
Anti-human CD45 PTPRC Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00555-Biotin | BosterBio | 100µg | 226 EUR |
Anti-Human CD45RO PTPRC Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC00555-FITC-1 | BosterBio | 25 Tests, 100 Tests, 200 Tests | 208 EUR |
Anti-human CD45 PTPRC Monoclonal Antibody PerCP Conjugated, Flow Validated |
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FC00555-PerCP | BosterBio | 25 Tests, 100 Tests, 200 Tests | 338 EUR |
Anti-human CD71 TFRC Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00591-Biotin | BosterBio | 100µg | 245 EUR |
Anti-human CD62L SELL Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC00652-Biotin | BosterBio | 100µg | 234 EUR |
Anti-human CD16 FCGR3A Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC01408-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-human CD32 FCGR2A Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC01450-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 268 EUR |
Anti-Human CD31 PECAM1 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC01513-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 235 EUR |
Anti-human CD90 THY1 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC01818-Biotin | BosterBio | 100µg | 267 EUR |
Anti-human CD20 MS4A1 Monoclonal Antibody Biotin Conjugated, Flow Validated |
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FC03780-Biotin | BosterBio | 100µg | 234 EUR |
Anti-human CD20 MS4A1 Monoclonal Antibody PerCP Conjugated, Flow Validated |
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FC03780-PerCP | BosterBio | 25 Tests, 100 Tests, 200 Tests | 338 EUR |
Anti-Human CD11a ITGAL Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC04466-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 231 EUR |
Anti-human CD57 B3GAT1 Monoclonal Antibody FITC Conjugated, Flow Validated |
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FC09548-FITC | BosterBio | 25 Tests, 100 Tests, 200 Tests | 245 EUR |
Von diesen wurde bestätigt, dass vier Klone (FHVH1–4) an eine andere vom Menschen stammende CD5+-Zelllinie (CCRF-CEM; Abbildung 1C) binden. Um ihre Bindungsaffinitäten zum CD5-Antigen weiter zu messen, wurden diese VH-Klone zusammen mit H65 im Format VH/scFv-Human-Immunglobulin-Fc-Fragment (hFc) oder VH/scFv-Kaninchen-Immunglobulin-Fc-Fragment (rFc) exprimiert und gereinigt. Die Bindungsaffinität wurde unter Verwendung des Octet96e-Systems bestimmt. FHVH1 (KD = 1,63 × 10 –9 M), FHVH2 (KD = 8,32 × 10 –9 M) und FHVH3 (KD = 1,39 × 10 –9 M) zeigten im Vergleich zu H65 ( KD = 6,53 × 10 –10 M), obwohl FHVH4 (KD = 2,47 × 10 –8 M) eine wesentlich niedrigere Affinität als die obigen Klone aufwies.