23 Jul Macrophagocyte transfer inhibition factor monoclonal antibody and method for making same
Abstract
The invention discloses a preparing method of monoclonal antibody (hMIF) to against human macrophage transferring inhibitor and getting method of amino acid sequence in biological medicine engineering technical domain, which comprises the following steps: augmenting out of hMIF gene from human breast cancer cell strain MDA-MB453 and human breast cancer tissue through reverse transcript polyose chain type reaction; constructing pET-11b/hMIF restructuring plasmid; expressing and purifying on bacillus coli BL21(DE3); immunizing Balb/c mouse; getting splenocyte of mouse and anti-hMIF monoclonal antibody 2D10 merged with SP2/0 cell through hybridomas technology; proceeding antibody subclass appraise and specificity test of antibody; making the secreting monoclonal antibody from 2D10 belong to IgG; setting the light chain as K type; setting the amino acid sequence of variable area of light chain as CRASGRNNAWYKGADGWYKGKAKRYAASSSGVSRSGSG SGTTTSSDATYYCHNDAVYSYGTKVKR; making the amino acid sequence of variable area as VWSGGSTVKG GRSCGYYASGTSTYSMGMNWVRAGKGWVSSSTSVDTYYYADSVKGRTSRDNAKNSYDY DVSRADTAVYYCARDGASSGWGVDATGSRVSCYWGGTVSSWMSAT. This anti-hMIF monoclonal antibody provides effective criterion for further transformation of human source, which can lay the groundwork for treating whole body inflammation in clinical therapeutics.
Year: 2007
Country: CN
Doc No: 101041820