Detection Of Anti-HBV PreS1(94-117) Antibody Via A Peptide Combination Of Multiple Genotypes

Chronic hepatitis B still imposes a threat to the health of Chinesecitizens with high prevalence. Although interferon has been expected asan effective treatment for chronic hepatitis B, its uncertain patientresponse, together with its high cost and possible side effects, has limitedits wide application, particularly in China.Previous studies in Europe and China have shown that the presenceof an antibody recognizing a specific peptide of amino acid94-117(P94)in the PreS1protein of hepatitis B virus (HBV) can be used as apre-treatment indicator for interferon response.However, the effectivenessof this biomarker needs to be improved to become a practical tool forclinical prediction of interferen response.Herein, we have developed a nature-mimiching peptide combinationapproach based on existing work to detect anti-P94antibodies in patientsera.We first screened HBV DNA sequences reported in GenBank andchose two P94peptide sequences representing most likely sequences inChinese patients. Then these peptides were used as coating antigen, eitherin combination or by each one itself, in enzyme-linked immunosorbentassay (ELISA). It has been found the method of peptide combination canindeed achieve more reliable results in patients of diversified HBVgenotypes. We also used P94petitdes coupled tokeyhole limpethemocyanin (KLH) to enhance ELISA signal and obtained good antigensspecific for the detection of anti-P94antibodies in Chinese hepatitis Bpatients. A scalable ELISA process for P94antibody detection has also been established.