[Preparation, characterization and application of monoclonal antibodies to Fc fragment of Ig fusion protein].
OBJECTIVE
To prepare and characterize mAb to Fc fragment of Ig fusion protein, and to establish sandwich ELISA for detecting Ig fusion proteins and affinity chromatography method for Ig fusion protein purification.


METHODS
hBCMA-Ig was used as antigen to immune BALB/c mice. The lymphocyte hybridoma technique was used to establish hybridoma cell lines stably secreting anti-Fc mAb. ELISA was employed to detect the isotype, epitopes, and species specificity of mAbs. Western blot was used to assess the reactivity between anti-Fc mAbs and denatured Ig fusion protein. LAIR1-Ig fusion protein was purified through affinity column anti-Fc mAb cross-linked to Sepharose 4B.


RESULTS
Seven hybridoma cell lines(FMUFc1-FMUFc7) were acquired. A sandwich ELISA was successfully established using FMUFc4 as coating mAb and FMUFc5 as enzyme-labeled mAb. FMUFc6 could be used for Western blot. LAIR1-Ig fusion protein was effectively purified through FMUFc5-Sepharose affinity chromatography column.


CONCLUSIONS
mAb against Fc fragment of Ig fusion protein has been prepared successfully, and methods to detect and purify Ig fusion protein are established. These mAbs provide useful tool for further application of Ig fusion protein.
Hu
Yi
Y
Deparment of Immunology, Fourth Military Medical University, Xi'an 710032, China.
Liu
Xue-Song
XS
Zhu
Yong
Y
Liu
Ying
Y
Xue
Xiao-Guang
XG
Xue
Jiang-Nan
JN
Jin
Bo-Quan
BQ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000276:immunology; D007141:Immunoglobulin Fc Fragments; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D007141:Immunoglobulin Fc Fragments