[Species-specific monoclonal antibodies against the major outer membrane protein (MOMP) of Chlamydia trachomatis].
The synthesized one quarter N-terminal MOMP of C. trachomatis was used for primary immunization of three male BALB/c mice (8 weeks of age), and the boost with C. trachomatis L1/440/Bu elementary bodies (EBs) was followed on day 14. Spleen cells from one mouse with good response of immunization were fused with murine myeloma NS-1 cells on day 24. The hybrid cell suspension was seeded into the wells of 96-well microtest plates which contained macrophage feeder layers. Anti-chlamydial antibodies in culture fluids were screened by ELISA with 1/4 MOMP & L1 EBs coated 96-well trays. Positive wells were cloned by limiting dilution. Four clones which secreted immunoglobulin G1 & G2a class were obtained after elimination of those clones that produced antibodies to C. psittaci strain EAE, C. pneumoniae strain ATCC VR1310 and uninfected BGMK cells. In micro-IF test, we found that the all four clones of MAbs reacted with our laboratory prepared L1, L2, A, B, C, E EBs, L2 tissue culture inclusions, as well as the EBs of all 15 standard serovars of C. trachomatis. The titers of their ascites were more than 1:12,800 in micro-IF test. It was shown that the four clones of MAbs reacted predominantly with 40,000 MOMP of C. trachomatis L1 in Western blot.
Ni
A
A
PUMC Hospital, CAMS, Beijing.
Everson
S
S
Li
Y
Y
Ward
M E
ME
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology*; D002692:Chlamydia trachomatis; D006825:Hybridomas / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018272:Porins; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D018272:Porins; C071677:omp1 protein, Chlamydia trachomatis