Development and identification of monoclonal antibodies against meso-Tetra (alpha,alpha,alpha,alpha,-O-phenylacetamide benzene) porphyrin.
The small molecule meso-Tetra (alpha,alpha,alpha,alpha-o-phenylacetamide benzene) porphyrin was synthesized through the condensation of o-nitrobenzaldehyde and pyrrole followed by reduction of the meso-tetra (o-nitrophenyl) porphyrin. The small molecule, without carrier, was used as complete antigen to immunize BALB/ C mice. Spleen cells producing high titer antibody were removed and fused with myeloma cells of SP2/0 origin. Using a conventional immunization protocol, stable murine monoclonal antibodies (MAbs) producing cell lines to meso-Tetra (alpha,alpha,alpha,alpha-o-phenylacetamide benzene) porphyrin 1F2 were obtained. Subclass determination showed that the clones produce IgG2a types of MAbs. The analytical results of HPLC and MALDI/TOFMS suggest that the purity of MAb 1F2 is 100%, and MAb 1F2 has a relative molecular weight of 156678.8 Da. Our results demonstrated that small molecule meso-Tetra (alpha,alpha,alpha,alpha-o-phenylacetamide benzene) porphyrin, as semiantigen without carrier, can elicit the formation of MAbs.
Wang
Fengyang
F
College of Life Science and Agriculture, Hainan University, Haikou, China. fywang68@yahoo.com
Huang
Xueying
X
Du
Li
L
Li
Weiguo
W
Qi
Chao
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D044482:Benzeneacetamides / Q000138:chemical synthesis / Q000276:immunology*; D002851:Chromatography, High Pressure Liquid; D007074:Immunoglobulin G / Q000096:biosynthesis*; D051379:Mice; D008807:Mice, Inbred BALB C; D011166:Porphyrins / Q000138:chemical synthesis / Q000276:immunology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000911:Antibodies, Monoclonal; D044482:Benzeneacetamides; D007074:Immunoglobulin G; D011166:Porphyrins; C522163:tetrakis(alpha,alpha,alpha,alpha-O-phenylacetamide benzene)porphryin