Application of multiple antigen peptide immunomagnetic beads in single-epitope antibody preparation

To prepare specific single-epitope antibody using immunomagnetic beads coated with multiple antigen peptides(MAP). 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 coat...

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Published inXi bao yu fen zi mian yi xue za zhi Vol. 23; no. 6; p. 572
Main Authors Xu, Xiao-jie, Zhang, Wen-hong, Kang, Lei, Han, Feng-chan, Yan, Zhen, Yan, Xiao-jun
Format Journal Article
LanguageChinese
Published China 01.06.2007
Subjects
Animals
Antibodies - immunology
Antibody Specificity
Electrophoresis, Polyacrylamide Gel
Epitope Mapping
Epitopes - immunology
Immunomagnetic Separation - methods
Mice
Mice, Inbred BALB C
Microspheres
Peptides - immunology
Rabbits
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Summary:To prepare specific single-epitope antibody using immunomagnetic beads coated with multiple antigen peptides(MAP). 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. 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%. 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
ISSN:1007-8738