Preparation And Preliminary Characterization Of Bispecific Antibody Against Anti-human Laryngeal Carcinoma And Anti-VEGF

Laryngocarcinoma is of high frequency in North China and its generation,regeneration&metastasis threaten lives of people.There are three difficulties in therapy of tumour for a long time:â‘ radiation&chemotherapy as the way of routin escheme have low distinctive to other tissues;â‘¡tumour cells become more and more drug resistant ;â‘¢very small metastasis of tumour. Immunotherapy is a new way to solve these problems.Using BispecificAntibody(BsAb) has been hot spot of immunotherapy for it can target tumour cells and have good prospects to clinical application.BispecificAntibody(BsAb) is monovalent in function but dualvalent in chemical structure.BsAb has more advantage than Monocolonal Antibodies(McAb).We used hybird-hyirdoma technique to prepare BsAb,that is to refuse two kinds of hyirdoma cell together,and it can secrete Ig with two kinds of binding specific function of the two McAbs.This special kind of Ig is what we were going to prepare. BsAb has been accepted as a new way of immunization therapyby by people.It can distinguish and combine with tumour cells,making tumour cells lose the ability of secreteing factors.Unlimited and invadable growth of malignant tumour rely on continue vessel generation.Many kinds of tumours can deliver VEGF much higher than other tissues, normal tissues deliver little,and the delivery of VEGF has much relationship with regeneration, metastasis&prognosis of tumours.Studies of in&out of the County has shown that with progressive rise of VEGF,it can promote new growth vessels,and in turn it promote the growth of laryngocarcinoma.With promotion of growth of new vessels and tumours,VEGF has been obviously related to periodization and differentiation of tumours. From the mechanism that the growth and metastasis of tumour depends on vessel growth,so any work which focus on restrain vessel growth will restrain tumour growth effectivel. For vessel growth depends on the irritatition of growthfactor ,anti-growthfactor is the key link of anti-tumour.One kind o tumour can secrete many kinds of vessel generation factor,one vessel generation factor can also been secreted by many kinds of tumours, as so to find out a kind of special vessel generation factor which can been distinguished by most of the tumours is of great value,and VEGF is this kind of one. So using antibody against VEGF will do great effect in tumour therapy and it will be a new way of anti-tumour to restrain physiological function of VEGF .The BsAb which we have prepared can not only combine with blood vessel endothelium cells of laryngocarcinoma,killing and restraining generation of vessel,restraining factors dischargement,but also can bind laryngocarcinoma cells distinctively,activating effect cells around tumour tissue,killing target cells,curative effect is more high and accurate,better than McAb.Through preparation of anti-human laryngeal carcinoma/anti-VEGF bispecific antibodies,we aim to findout a new way to cure laryngeal carcinoma.We use splenocytes of Balb/c mice which had been immunized with the antigen of human laryngeal carcinoma or VEGF,were used to fuse with myeloma cells of NS-1 mice separately.Hybird-hyirdoma technique was used to prepare BsAb and it was analysed and characterized with ELISA et al.After screening, 5 stains of hybirdomas which could secrete McAb of anti-laryngeal carcinoma or anti-VEGF each and one stain of BsAb were obstained.The BsAb was of high purity and specificity.In all,we have prepared BsAb successfully and it will be great helpful to laryngeal carcinoma therapy.