Two new murine monoclonal antibodies rised against human IgG
Many pathological conditions are accompanied with changes in the concentration of the total IgG or some of its fraction. For this reason there is great interest in the production of reagents specific for IgG. In this paper, the binding characteristics of two new murine monoclonal antibod...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Serbian Chemical Society
2001-01-01
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| Series: | Journal of the Serbian Chemical Society |
| Subjects: | |
| Online Access: | http://www.doiserbia.nb.rs/img/doi/0352-5139/2001/0352-51390105323P.pdf |
| Summary: | Many pathological conditions are accompanied with changes in the
concentration of the total IgG or some of its fraction. For this reason
there is great interest in the production of reagents specific for IgG. In
this paper, the binding characteristics of two new murine monoclonal
antibodies (MoAb), assigned MoAb 15 and MoAb 22, are reported. These MoAbs
were produced by hybridoma technology. By performing ELISAs and Western
blots analyzes, it was demonstrated that both MoAbs interact specifically
with human IgG. Cross reactivity with other sera proteins was not observed.
In order to precisely localize the epitopes recognized by MoAb 15 and MoAb
22, the Western blots interactions of these MoAbs with electrophoreticaly
separated IgG-fragments, obtained by the action of proteolytic enzymes
(papain, pepsin, trypsin), were analyzed. According to the results of these
experiments, both MoAbs interacted with epitopes in the C 3 domain. The
affinity constants, calculated from Scatchard plots of binding ofMoAb15
and MoAb22 to human IgG, wereKa15 = 1.71 106M-1 andKa22 = 2.15 109M-1.
According to all these findings,MoAb 15 and MoAb 22 could be used in
standard immunochemical techniques. However, the experiments showed that
both MoAbs had bad immunoprecipitating properties. In solid phase techniques
(ELISAs, Western blot, dot-blot, etc.), their application gave excellent
results that highly recommended them for use in these types of analyzes. |
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| ISSN: | 0352-5139 1820-7421 |