[Generation and identification of phage engineering antibodies library against hamster prion protein].
OBJECTIVE
Generation and Identification of Phage Engineering Antibodies Library against Hamster Prion Protein.


METHODS
Fab antibodies were identified and confirmed. BALB/c mice were immuned with PrP proteins. After the third immunization, the total RNA was extracted from the mice spleens. The genes of heavy Fd Fragments and light chain of antibodies amplified by a series of specific primers of human IgG Fab fragment were cloned into phagemid vector pComb3.


RESULTS
The combinatorial Fab library were constructed successfully and the cloning efficiencies both of light chain and Fd fragments were about near 10(6). The Fab library were panned by four cycles and screened with purified haPrP23-231 antigen on microtiter plates. 12 mAbs were isolated after four cycles of panning, five of which were sequenced and resulted sequence data were analyzed by alignment with GenBank immunoglobulin genes. Two strain of new heavy and light chain genes of Fab antibodies were identified and confirmed.


CONCLUSIONS
The research in this article will provide foundation for study of diagnosis and therapy of prion.
Huang
Yin-Xia
YX
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
Han
Jun
J
Dong
Chen-Fang
CF
Sun
Li
L
Gao
Chen
C
Wang
Xiao-Fang
XF
Han
Lu
L
Zhou
Wei
W
Zhang
Bao-Yun
BY
Jiang
Hui-Ying
HY
Liang
Mi-Fang
MF
Dong
Xiao-Ping
XP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D006224:Cricetinae; D005260:Female; D006367:HeLa Cells; D006801:Humans; D007114:Immunization; D007140:Immunoglobulin Fab Fragments / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D019151:Peptide Library; D011328:Prions / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D011328:Prions