Development And Characterization Of Recombinant Therapeutic Human Monoclonal Antibody That Neutralize TGF-?1

The role of TGF-? in tumor development and progression is complex.Genetic mutations that disrupt the antiproliferative signaling effects of TGF-? play a key role in the process of malignant transformation for many types of tumors.Paradoxically,this loss of sensitivity to TGF-?'s inhibitory actions often leads to TGF-? over-expression by the tumor cells or by normal cells that are recruited to the tumor microenvironment.Elevated concentrations of TGF-? in the tumor microenvironment have been shown to facilitate tumor growth and metastasis.Therefore,selective inhibition of excessive TGF-? is being pursued as a therapeutic strategy for hTGF-? related disease.Numerous published studies have provided evidence that inhibition of TGF-? using antibodies,soluble receptors and small molecule inhibitors of TGF-? signal transduction can have beneficial effects in murine models of cancer.In this work,we aimed to discovery a therapeutic leads of antibody with independent intellectual property rights which binding to hTGF-? 1 and yet demonstrated its ability to suppress tumor growth;which were based on the these two antibodies,one is come from the hybridoma sources named as 1D11,which is belongs to the R&D company;and the other is a humanized antibody LY-2382770 from the Eli Lilly company.And finally to establishment a bifunctional antibody with PD-1 antibody or treatments with combination therapy.On study of this two antibodies,we reveal that the two antibodies share the same binding epitope but the 1D11 antibody have a significantly lower avidity and biological activities than the LY-2382770 antibody;the latter one also exists the shortage of the thermodynamic stability.After considering all these factors,we decide to select the LY-2382770 antibody as an optimal molecule for antibody engineering.As for the antibody Vh gene is dominantly involved in the determination of the antigen specificity,to minimize the deteriorative influences on the TGF-? specificity of Lilly hIgG4 antibody LY-2382770,we aimed to achieve the thermostability modification of Lilly hIgG4 antibody LY-2382770 using Kappa-chain shuffling library and select LY-2382770 antibody variants.The experimental results show that the use of Kappa chain shuffling can effectively improve the antibody's thermal stability and expression yields,ameliorate the aggregation of antibodies,and does not have a significant impact on the affinity of the antibody itself.The use of affinity maturation methods can compensate for the decrease in affinity,and the combined use of these two methods can speed up the pace of research and development of lead molecules and development efficiency.