[Preparation and characterization of monoclonal antibodies against human laminin].
OBJECTIVE
To prepare monoclonal antibody(mAb) against humen laminin(LN) and identify their properties.


METHODS
BALB/c mice were immunized with humen LN, and then mAb was prepared by hybridoma technique. The titres of mAbs in ascetic fluid and relative affinity of mAb were determined by indirect ELISA test. Ig subgroup, specificity and combine dot of mAb were analysed by ELISA.


RESULTS
4 hybridoma cell lines 2A3, 2C6, 3G7 and 4H2 secreting anti-humen LN mAb were obtained. Their Ig subgroup belonged to IgG1, light chains were type kappa, The titres of 4 mAbs in ascitic fluid were 3.6 x 10(4)-2.1 x 10(6). Relative affinity of mAb 2C6 was above 10(12) and relative affinity of other 3 mAbs 2A3, 3G7 and 4H2 was 10(6), 10(8) and 10(8). Epitope recognized by 2 mAb(2A3 and 4H2) differed from the that by 2C6 and 3G7 another.


CONCLUSIONS
4 mAbs against humen LN has been prepared successfully and possess higher titer, specificity and relative affinity. Preparation of anti-human LN mAbs provide a useful reagent for further studying the role of LN in some diseases.
Yao
Min-jie
MJ
Jilin Military Medical College Affiliated, Fourth Military Medical University of Chinese PLA, Jilin 132013, China.
Huang
Han-chao
HC
You
Lian-xiu
LX
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis / Q000276:immunology; D007797:Laminin / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007145:Immunoglobulin kappa-Chains; D007797:Laminin