[Protein array technology applied in high throughput monoclonal antibody generation].
To reduce the huge labor-cost in the screening in traditional monoclonal antibody generation, We established a new system for monoclonal antibody generation integrating with protein array. BALB/c mice were immunized by eight recombinant proteins respectively, and the positive hybridoma cells were obtained by cell fusion and ELISA screening. All the eight kinds of positive hybridoma cells were mixed, cloned, screened by protein array, and definite dilution cloned.


RESULTS
175 single cell clones were obtained by complex cloning, and 119 of those were positive clones. Then 8 positive cell lines were generated by the following 2 rounds definite dilution cloning. By comparing with the traditional method, we got 8 monoclonal antibodies using the combined protein array screening and multiplex cloning method in 1 cycle, and fewer amounts of antigens were used. As a result, the combined protein array and multiplex cloning method could be used as an economical, rapid and simple tool applying in high throughput monoclonal antibody generation.
Song
Kai
K
National Engineering Center for Biochip at Shanghai, Shanghai Biochip Co., Ltd, Shanghai 201203, China.
Ye
Sai
S
Zhou
Jia-Jing
JJ
Peng
Hai-Lin
HL
Wang
Sheng-Nian
SN
Wei
Ling
L
Xiao
Hua-Sheng
HS
Zhao
Guo-Ping
GP
Zhang
Qing-Hua
QH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology* / Q000302:isolation & purification; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D040081:Protein Array Analysis
D000911:Antibodies, Monoclonal