致仔猪过敏性大豆抗原蛋白β-conglycinin单克隆抗体的制备与鉴定

研究以人工合成的对应于β-conglycinin分子α'亚基上的一段抗原表位肽(RPQHPER,78-84α')作为半抗原,与载体蛋白(OVA)交联后,作为免疫原制备了致仔猪过敏性大豆抗原蛋白β-conglycinin的单克隆抗体(Mab 6G4),并应用酶联免疫吸附等方法进行鉴定。结果表明,制备的单抗Mab 6G4属于IgG1;该抗体能与大豆β-conglycinin特异性结合,IC50为4.7 ng/mL,并对β-conglycinin表现出较强的亲和力,亲和常数达6.9×109 L/mol。因此,制备的单克隆抗体Mab 6G4为分析大豆制品中β-conglycinin的含量及探讨β-conglycinin对仔猪的致敏机理提供了一个潜在的分析工具。

Preparation and Characterization of Monoclonal Antibodies against Soybean β-conglycinin, a Hypersensitive Allergen to Piglets

This experiment was conducted to prepare monoclonal antibodies against soybean β-conglycinin using a conjugate of chicken ovalbumin and a synthetic peptide that corresponded to one epitope sequence,the amino-acid residues 78-84 present in theα' subunit of β-conglycinin,as the immunogen.The developed Mab,named as 6G4,was identified as being of the IgG1 isotype and showed high specificity to β-conglycinin with an IC50 of 4.7 ng/mL and a Ka of 6.9×109 L/mol by enzyme-linked immunosorbent assay.Therefore,the Mab(6G4) could provide a potential tool for quantification of β-conglycinin in soybean and researches on the hypersensitivity mechanism of β-conglycinin in piglets.