[Preparation and characterization of monoclonal antibodies against recombinant human NDPK-A].
OBJECTIVE
To prepare monoclonal antibodies (mAb) against recombinant human nucleoside diphosphate kinase-A(NDPK-A) and characterize their properties.


METHODS
BALB/c mice were immunized with rhNDPK-A, and mAb was prepared by hybridoma technique. Ig subclass and specificity was analysed by double immunodiffusion and western blot respectively. The titres of mAbs in ascitic fluid, relative affinity and epitopes recognized by mAbs were determined by indirect ELISA.


RESULTS
6 hybridoma cell lines secreting anti-rhNDPK-A mAbs were obtained. Their Ig subclass belonged to IgG1. The titers of 6 mAbs in ascitic fluid were 1x10(-4)-5x10(-6). Relative affinity of mAbs were 4.5x10(-9)-2.8x10(-10) mol/L. They recognized 3 different epitopes on rhNDPK-A molecule.


CONCLUSIONS
6 mAbs against rhNDPK-A have been prepared successfully which provide useful reagent for clinical diagnosis and further research.
Li
Xue-ling
XL
Biomedicine Research & Development Center of Jinan University, Guangzhou 510632, China.
Wang
Yi-fei
YF
Xiong
Sheng
S
Zhang
Mei-ying
MY
Li
Bing
B
Li
Jiu-xiang
JX
Wang
Hong
H
Zheng
Zuo-ya
ZY
Wang
Cong-zhu
CZ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D054778:NM23 Nucleoside Diphosphate Kinases; D009701:Nucleoside-Diphosphate Kinase; D011506:Proteins / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D054778:NM23 Nucleoside Diphosphate Kinases; D011506:Proteins; D011994:Recombinant Proteins; C517939:Nme1 protein, mouse; D009701:Nucleoside-Diphosphate Kinase