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Updated: Jul 14, 2026
Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
Published on: March 24, 2017
[Application of multiple antigen peptide immunomagnetic beads in single-epitope antibody preparation]
Xiao-jie Xu1, Wen-hong Zhang, Lei Kang
1Institute of Genetic Diagnosis, Fourth Military Medical University, Xi'an 710032, China.
Aim:
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%.
Conclusion:
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.
