Study On Stability Of 4E10-MAP4 Peptide Vaccine Preparation

AIDS(Acquired Immune Deficiency Syndrome),infectious disease is harmful to human survival and development,the world now has about 1% of the total population of people infected with HIV,the AIDS epidemic Chinese is the world's worst countries,AIDS is urgent.Since thirty years ago to find and confirm the HIV virus,after many efforts,researchers have made great progress,the cocktail therapy for the treatment of AIDS patients can greatly prolong the period of their disease,and help them to rebuild the immune system,but this method does not eliminate fundamentally the AIDS virus.Therefore,one of the best ways to control the spread of HIV is undoubtedly a safe and effective vaccine.However,none of the currently developed proteins,viral vectors,nucleic acids and polypeptide vaccines can be clinically proven to be effective.And some vaccines have exacerbated HIV infections.Safe broad-spectrum neutralizing antibodies are expected in the HIV vaccine.The HIV envelope protein gp41 MPER,is the target of broad-spectrum neutralizing antibodies(4E10,2F5,10E8 and Z13),HIV virus infected cells,MPER can produce a series of conformational changes,brief exposure of the epitope can be recognized by the immune system,antibody and identified epitopes binding.The inhibition of MPER area further conformational change,terminate the infection of HIV virus on the cell.In 2013,the relevant research shows that using MAPs(branched peptide form)broad-spectrum neutralizing antibody 4E10 presenting MPER epitope,and immune to guinea pigs after conditioning,supplemented with oil adjuvant,can successfully induce epitope specific antibody response,pseudotyped virusneutralization test results showed that 4E10-MAP4 induced by broad-spectrum the neutralizing activity,and subsequent related research,initially established a 4E10-MAP4 formulation,and the optimal immunization program.According to the quality of the early 4E10 MAP4 immunogen,this paper studies the stability of 4E10-MAP4 clinical preparation according to the relevant provisions of the Chinese pharmacopoeia.According to the 2015 edition of "Chinese Pharmacopoeia"(four guiding principles of the 9001 drug substances and drug stability test guidelines)in the long-term test and accelerated test development time,Long test time is 0 months,3 months,6 months,12 months,18 months and 36 months.Accelerated test for study time was zeroth days,7 days,14 days,21 days,28 days,the effects of temperature is 20~25? and 36±1?.The free peptide content by UV spectrophotometry and HPLC determination of medium.Determination of polypeptide content in preparations by MS and HPLC.To calibrate the effective preparation of antigenicity by Western blotting and ELISA.That is always in line with standard antigenic preparation.Through the measurement and calculation of animal immune positive rate,prove the efficacy of preparations.The physical and chemical properties of the preparation were determined by visual inspection,measurement and pH measurement.The test results show that the content of polypeptide and preparation quality does not change with time,physical and chemical properties meet the requirements of Chinese pharmacopoeia.Next,through the 20~25?,36±1? accelerated test,the appearance of the product,the amount of installed,pH were measured,the test results show that the preparation does not because of short-term temperature rise,physical and chemical properties have changed.In summary,the polypeptide AIDS vaccine developed by the laboratoryaccording to the listed sales package at 2~8? conditions preserved for36 months,the indicators are in line with the provisions and the property is stable,so the polypeptide vaccine is valid for 36 months.The preparation can be packaged,transported and stored at 20~25? and 36±1?,and the product quality can not be affected as long as the time is not more than 28 days.