Khosraviani M, Nunoya T, Matsumoto M
College of Veterinary Medicine, Oregon State University, Corvallis 97331.
Avian Dis. 1990 Jan-Mar;34(1):163-73.
Four monoclonal antibodies (MAbs) were developed against serotype 3:A, P-1059 strain of Pasteurella multocida. Enzyme-linked immunosorbent assays were used to screen those hybridomas producing antibodies to either a surface protective (2.5 S) or lipopolysaccharide (LPS) antigen. MAbs 6EE11, D7H10, E11E3, and C11H2 were positive against 2.5 S antigen, and two of them, E11E3 and C11H2, were positive for the LPS antigen. MAbs 6EE11 and D7H10 reacted with a major protein band of molecular weight of 35,500, whereas E11E3 and C11H2 recognized a band with a molecular weight of 12,500 of the 2.5 S antigen. Treatment of the 2.5 S antigen with periodic acid abolished epitopes reacting with E11E3 but not with 6EE11. MAb 6EE11 did not recognize any band in Western blot after proteinase K treatment of the 2.5 S antigen, whereas antibody activity of E11E3 did not change. MAb 6EE11 reacted with serotypes 3, 4, 9, 10, 11, 12, and with M-9 strains in the immunofluorescence test. MAb E11E3 was positive only with serotype 3 or 10 strains, excluding M-9 strain. Electron microscopic studies with P-1059 strain indicated that antigens binding to 6EE11 and/or E11E3 were present in the capsule.
针对多杀性巴氏杆菌血清型3:A、P - 1059菌株研制了四种单克隆抗体（MAb）。采用酶联免疫吸附测定法筛选那些产生针对表面保护性（2.5S）或脂多糖（LPS）抗原抗体的杂交瘤。单克隆抗体6EE11、D7H10、E11E3和C11H2对2.5S抗原呈阳性反应，其中E11E3和C11H2两种对LPS抗原呈阳性反应。单克隆抗体6EE11和D7H10与一条分子量为35,500的主要蛋白带发生反应，而E11E3和C11H2识别2.5S抗原中一条分子量为12,500的条带。用高碘酸处理2.5S抗原消除了与E11E3反应的表位，但未消除与6EE11反应的表位。用蛋白酶K处理2.5S抗原后，单克隆抗体6EE11在蛋白质印迹法中未识别出任何条带，而E11E3的抗体活性未改变。在免疫荧光试验中，单克隆抗体6EE11与血清型3、4、9、10、11、12以及M - 9菌株发生反应。单克隆抗体E11E3仅对血清型3或10菌株呈阳性，不包括M - 9菌株。对P - 1059菌株进行的电子显微镜研究表明，与6EE11和/或E11E3结合的抗原存在于荚膜中。
