禽源多杀性巴氏杆菌(A:1)荚膜保护性抗原提纯及单克隆抗体细胞系建立

Purification of a Protective Capsule Antigen from Pasteurella Multocida (5: A) of Pcultry Origin and Establishment of the McAb Hybriodomas to It

用Sep(?)adex G-200分之筛层析、DEAE-cellulose离子交换层析相结合,将禽源多杀性巴氏杆菌(A:1)荚膜抗原粗提物(CE)分为P1.P2两种成分,并经免疫保护试验证明,荚膜中含有保护性蛋白成份P1。P2分子量为129 100,且至少含有分子量分别为46700、42600和39800的3种蛋白亚基。用CE免疫BALB/c小鼠,建立了抗P1的单克隆抗体细胞系1B1和2B7,它们只与多杀性巴氏杆菌反应,而不与其他细菌交叉。

The capsule crude extracts (CE) of Pasteurella multocida (5: A) could be di-vided into P1 and P2 fractions using the mechanical-physical purification combined with Sephadex G-200 gel filtration chromatography and DEAE-cellulose ion exchange chromatography. The protective protein of the capsule was detected only in P1 with the im-mune protective test. The molecular weight of P1 is 129 100 and it at least consists of three protein subunits whose molecular weight are 46 700, 42 600 and 39 800, respectively. Two hybridomas secreting monoclonal antibodies against P1 were established by means of the fu-sion between mouse myeloma cells SP2/0 and the spleen cells from BALB/c mouse immu-nized with CE. McAbs, 1B and 2B7,responded only to Pasteurella multocida and did not to other bacteria.