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中国生物工程杂志

China Biotechnology
China Biotechnology  2009, Vol. 29 Issue (05): 44-49    DOI:
    
Expression and Analysis of Humanized Anti-HER2 Monoclonal Antibody in Kluyveromyces lactis
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Abstract  Objective: To express humanized anti-HER2 monoclonal antibody in Kluyveromyces lactis. Methods: The heavy chain and light chain genes of humanized anti-HER2 monoclonal antibody were amplified using PCR and inserted into yeast expression vector pYES 2/ochI and pPICZαA/ura3,respectively. The restriction enzyme analysis and DNA sequence direction confirmed that the inserted fragment of clone was the right humanized anti-HER2 monoclonal antibody coding sequence. The recombinant vectors were transformed into the host cell Kluyveromyces lactis(Δura3)and induced with galactose. The expression level and the potential of antigen combination were detected by Indirect-ELASA and Western blot. Results: The recombinant expression plasmids pYES 2/ochI+αL and pPICZαA/ura3+αH were constructed, the expression level of humanized anti-HER2 monoclonal antibody in flask reached (120±20)mg/L. The purified product was analyzed by reduced and nonreduced SDS-PAGE. The result showed that the light chain and heavy chain of antibody could assemble as the right antibody structure through disulfide bonds spontaneously. The antibody expressed in K.lactis also had the potential ability of specific combination with its antigen. Conclusion: The humanized anti-HER2 monoclonal antibody which had high potential of antigen combination was effectively expressed in Kluyveromyces lactis.

Key wordsKluyveromyces lactis;High Secreted Expression;Anti-HER2 Monoclonal Antibody     
Received: 24 November 2008      Published: 06 May 2009
Cite this article:

HONG Li-Na- Liu-Bei- Gong-Xin- Chang-Shao-Gong- Wang-Ling-Xue- Song-Miao- Xu-Wei- Tun-Jun. Expression and Analysis of Humanized Anti-HER2 Monoclonal Antibody in Kluyveromyces lactis. China Biotechnology, 2009, 29(05): 44-49.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2009/V29/I05/44

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