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目的通过竞争性ELISA方法明确牙龈卟啉单胞菌血凝素2(Porphyromonas gingivalis hemagglutinin-2,Pg HA-2)与氯化血红素结合的多肽位点,为牙周病保护性抗体的制备奠定基础。方法人工合成疑为Pg HA-2氯化血红素结合位点的多肽片段,制备抗Pg HA-2单克隆抗体(MAb QB),通过间接竞争性ELISA,进一步分析血红素特异性结合位点。结果 Pg HA-2氯化血红素结合位点的氨基酸序列为DGFPGDHYAVMISK。MAb QB可以抑制Pg HA-2与氯化血红素结合。结论明确了Pg HA-2氯化血红素结合位点的氨基酸序列,确定了Pg HA-2氯化血红素结合位点的位置。为今后Pg HA-2氯化血红素结合位点的鉴定、功能结构区分析和多肽疫苗的制备奠定基础。
OBJECTIVE To determine the binding site of Porphyromonas gingivalis hemagglutinin-2 (Pg HA-2) and hemin by competitive ELISA, and lay the foundation for the preparation of protective antibodies against periodontal disease basis. Methods Anti-Pg HA-2 monoclonal antibody (MAb QB) was prepared by artificial synthesis of the polypeptide fragment suspected to be the heme binding site of Pg HA-2. Heme-specific binding sites were further analyzed by indirect competitive ELISA. Results The amino acid sequence of Pg HA-2 hemin binding site was DGFPGDHYAVMISK. MAb QB inhibits the binding of Pg HA-2 to hemin. Conclusion The amino acid sequence of hemin binding site of Pg HA-2 was confirmed, and the position of hemin binding site of Pg HA-2 was determined. Pg HA-2 for the future identification of hemin binding sites, functional structure analysis and preparation of polypeptide vaccine laid the foundation.