Anti-p185<erbB2> human-mouse chimeric antibody lentivirus expression vector and construction method thereof

Abstract

The invention discloses an anti-p185<erbB2> human-mouse chimeric antibody lentivirus expression vector and a construction method thereof. Antihuman-p185<erbB2> mouse-derived monoclonal antibody variable region genes (vH and vL) and human-IgG1 constant region genes (gamma 1 and kappa) are obtained by amplification by PCR (polymerase chain reaction) process. The vH is spliced with the gamma 1 and the vL is spliced with the kappa by three-primer PCR process so as to obtain a chimeric heavy chain (H) gene and a chimeric light chain (L) gene. The chimeric heavy chain (H) gene and the chimeric light chain (L) gene are inserted onto an IRES (internal ribosome entry site) element of plasmid pVAX1 / IRES downstream and upstream respectively. H-IRES-L is cut off from the pVAX1 / H-IRES-L using endonucleases and is inserted into the lentivirus expression vector pWPI so as to construct a lentivirus expression pWPI / H-IRES-L. Corresponding enzyme digestion and sequencing appraisal show that the expression is consistent to that of the expected design. After transfection of cells 293T, the chimeric heavy chain (H) gene and the chimeric light chain (L) gene are co-expressed, and the chimeric antibody can be combined with p185<erbB2> molecular specificity. By the vector and the construction method, basis is laid for anti-p185<erbB2> engineering antibodies in the future.

Description

technical field [0001] The present invention relates to a lentiviral expression vector, especially an anti-p185 erbB2 Human-mouse chimeric antibody lentiviral expression vector and its construction method. Background technique [0002] c-erbB2 / HER2 / neu encoded product p 185 erbB2 It is a transmembrane glycoprotein with tyrosine kinase activity and belongs to the epidermal growth factor receptor (EGFR) family. Amplification of c-erbB2 / HER-2 / neu gene and p185 erbB2 Overexpression of the protein is seen in a variety of malignancies, including breast, ovarian, colon, endometrial, gastric, prostate, and lung adenocarcinomas. p185 erbB2 Overexpression indicates poor prognosis of tumors, such as metastasis, recurrence, easy drug resistance and short survival period. Therefore, p185 erbB2 As a tumor antigen, it is an ideal target for tumor therapy. In October 1998, the U.S. Food and Drug Administration (FDA) officially approved the introduction of an anti-p185 erbB2 Humanize...

AI Technical Summary

Problems solved by technology

However, imported antibody drugs are expensive, how to develop anti-p185 with independent intellectual property rights erbB2 Genetic engineering antibodies have very important practical significance for domestic patients, among which, the construction of expression vectors is particularly critical

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