[Application of multiple antigen peptide immunomagnetic beads in single-epitope antibody preparation].
OBJECTIVE
To prepare specific single-epitope antibody using immunomagnetic beads coated with multiple antigen peptides(MAP).


METHODS
Eight-branch MAP is synthesized by Fmoc SPPS. After immunizing the BALB/c mice with the MAP, polyclonal antiserum was obtained. Then, the immunomagnetic beads (IB) were prepared by coating with the MAP in covalent bond form and were used to purify the specific single-epitope antibody from the antiserum of immunized animals.


RESULTS
The titer of antiserum was 1:12,800, which showed the strong immunogenicity of MAP. The best coating concentration of MAP was 100 mg/L and the max coating efficiency was 79%. After purifying specific antibody from polyclonal antiserum using the IB, the obtained antibodies only interacted with one epitope rather than other epitopes. The purity of the antibody reached 95% and the mean recovery rate was 5.8%.


CONCLUSIONS
The beads coated with MAP can be used to quickly isolate specific single-epitope antibody which is similar to monoclonal antibody. This method provide a convenient shortcut in preparing small amount of antibody with high specificity.
Xu
Xiao-jie
XJ
Institute of Genetic Diagnosis, Fourth Military Medical University, Xi'an 710032, China.
Zhang
Wen-hong
WH
Kang
Lei
L
Han
Feng-chan
FC
Yan
Zhen
Z
Yan
Xiao-jun
XJ
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D004591:Electrophoresis, Polyacrylamide Gel; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology*; D018189:Immunomagnetic Separation / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D008863:Microspheres; D010455:Peptides / Q000276:immunology*; D011817:Rabbits
D000906:Antibodies; D000939:Epitopes; D010455:Peptides