Preparation and activity detection of monoclonal antibody against anti-CD3 ScFv / 中国医学科学院学报
Acta Academiae Medicinae Sinicae
;
(6): 354-359, 2008.
Artículo
en Chino
| WPRIM
| ID: wpr-270689
ABSTRACT
<p><b>OBJECTIVE</b>To prepare monoclonal antibody (McAb) against anti-CD3 ScFv for purifying and detecting serum anti-CD3 antibody concentration.</p><p><b>METHODS</b>McAb against anti-CD3 ScFv was prepared by hybridoma technique and used to prepare affinity chromatography column, which was used to purify anti-CD3 ScFv and Diabody [CD3 x Pgp] without E-tag. The binding activities of anti-CD3 ScFv, Diabody [CD3 x Pgp] without E-tag, and Diabody [CD3 x Pgp] purified by anti-CD3 affinity chromatography column or anti-E-tag affinity chromatography column against K562/A02 cell and Jurket cells were detected by fluorescence activated cell sorting (FACS) method. ELISA was used to identify the specificity of the McAb.</p><p><b>RESULTS</b>McAb against anti-CD3 ScFv specifically detected serum anti-CD3 ScFv without reacting with sera. The anti-CD3 ScFv purified by anti-CD3 affinity chromatography column and purified by anti-E-tag affinity chromatography column had the same specific binding activity with Jurkat cells. The positive binding rates of Diabody [CD3 x Pgp] without E-tag to K562/A02 and Jurkat cells were 89.87% and 83.95%, respectively. In the competitive binding experiments with K562/A02 and Jurkat cells, the binding rates of Diabody [CD3 x Pgp] without E-tag decreased to 56.30% and 43.78%, respectively.</p><p><b>CONCLUSION</b>The McAb against anti-CD3 ScFv prepared in our lab can be used to purify and detect serum anti-CD3 antibody concentration.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Línea Celular
/
Cromatografía de Afinidad
/
Complejo CD3
/
Células Jurkat
/
Células K562
/
Alergia e Inmunología
/
Hibridomas
/
Metabolismo
/
Anticuerpos Monoclonales
Tipo de estudio:
Estudio diagnóstico
Límite:
Humanos
Idioma:
Chino
Revista:
Acta Academiae Medicinae Sinicae
Año:
2008
Tipo del documento:
Artículo
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