[Purification and identification of pregnancy associated plasma protein A].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
; 27(8): 835-8, 2011 Aug.
Article
en Zh
| MEDLINE
| ID: mdl-21806876
ABSTRACT
AIM:
To purify pregnancy associated plasma protein A (PAPP-A) from the pregnancy serum using the methods of G200 gel filtration, reverse affinity chromatography and DEAE ion exchange.METHODS:
The male sera were purified by G200 gel filtration and injected into rabbits to obtain rabbit-anti-human antibodies. The antibodies were linked to CNBr-activated G25 gel to make the reverse affinity chromatography. The full-term pregnancy sera were collected and centrifuged, followed by G200 gel filtration, reverse affinity chromatography and DEAE ion exchange. ELISA and SDS-PAGE were performed to detect the activity and the purity of PAPP-A.RESULTS:
As detected by the double diffusion, the rabbit-anti-human antibodies were active in the dilution of 116, and were further used for purification of PAPP-A. The PAPP-A exited in the last peak with the pH3.0 elutriant by SPA, the first peak by G200 gel filtration, the peak with 0.1 mol/L PBS elutriant by the reverse affinity chromatography, and the peak with 0.45 mol/L NaCl elutriant by the DEAE ion exchange. The SDS-PAGE results indicated that the triple treatments of the pregnancy sera by G200 gel filtration, the reverse affinity chromatography and the DEAE ion exchange effectively enhanced purity of PAPP-A, with a single protein band shown by Western blot, whereas a single purification method produced dirty bands.CONCLUSION:
High purity PAPP-A has been obtained, ready for preparation of its monoclonal antibody. The study paves a way for development of PAPP-A based ELISA kits for clinical use.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Proteína Plasmática A Asociada al Embarazo
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
/
Risk_factors_studies
Límite:
Adult
/
Animals
/
Female
/
Humans
/
Male
/
Middle aged
/
Pregnancy
Idioma:
Zh
Revista:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
Asunto de la revista:
ALERGIA E IMUNOLOGIA
Año:
2011
Tipo del documento:
Article
País de afiliación:
China