Author/Editor		Fan, Xiao-Hui
Title		Priprava himernih protiteles proti človeškemu katepsinu B
Translated title		(Preparation of chimeric antibody against human vathepsin B)
Type		monografija
Place		Ljubljana
Publisher		Medicinska fakulteta
Publication year		2002
Volume		str. 114
Language		eng
Abstract		A mouse monoclonal antibody (MAb) against human cathepsin B which binds an epitope near the active site of cathepsin B has been prepared by our group at Jožef Stefan Institute (Aleš Premzl et al., submitted). This MAb is able to block cathepsin B activity and significantly impairs the invasion of ras-transformed MCF-10A neo T cells in virro. The aim of this work is to confirm the hypothesis that a chimeric monoclonal antibody raised against cathepsin B, capable of neutralizing its proteolytic activity, will enhance its possible clinical use in cancer therapy and in other diseases associated with uncontrolled over-expression of cathepsin B. One of the major problems for the therapeutic application of murine MAbs is the human anti-mouse antibody (HAMA) response. In order to reduce the HAMA response, recombinant DNA technology was used to construct a chimeric antibody containing constant human and variable murine regions. For the light chain, after amplification of VL fragment with primers L5V and L3V, the PCR product was subcloned into pUCSechCK, which contained human CK gene and a gene designed to provide the recombinant chimeric chains with a leader peptide required for secretion in extracellular medium. Finally, the chimeric light chain was cloned into the eukaryotic expression vector pcDNA3. For the heavy chain, after amplification of VH fragment with primers HSVB and H3V, it was subcloned into pUTSec vector, which also contained a gene for the leader peptide. Afterwards it was subcloned into pUC-hIgG1-Sal I vector containing human Cgamma1 gene. The chimeric heavy chain was also cloned into the vector pcDNA3. (Abstract truncated at 2000 characters).
Descriptors		CATHEPSIN B ANTIBODIES, MONOCLONAL CHIMERIC PROTEINS NEOPLASMS, EXPERIMENTAL PLASMIDS MICE ANTIBODIES, BACTERIAL TRANSFECTION POLYMERASE CHAIN REACTION SEQUENCE ANALYSIS, DNA