Ribosome Display Selection of a Murine IgG(1) Fab Binding Affibody Molecule Allowing Species Selective Recovery Of Monoclonal Antibodies
2011 (English)In: Molecular Biotechnology, ISSN 1073-6085, E-ISSN 1559-0305, Vol. 48, no 3, 263-276 p.Article in journal (Refereed) Published
Affinity reagents recognizing constant parts of antibody molecules are invaluable tools in immunotechnology applications, including purification, immobilization, and detection of immunoglobulins. In this article, murine IgG(1), the primary isotype of monoclonal antibodies (mAbs) was used as target for selection of novel binders from a combinatorial ribosome display (RD) library of 10(11) affibody molecules. Four rounds of selection using three different mouse IgG(1) mAbs as alternating targets resulted in the identification of binders with broad mIgG(1) recognition and dissociation constants (K (D)) in the low nanomolar to low micromolar range. For one of the binders, denoted Z(mab25), competition in binding to full length mIgG(1) by a streptococcal protein G (SPG) fragment and selective affinity capture of mouse IgG(1) Fab fragments after papain cleavage of a full mAb suggest that an epitope functionally overlapping with the SPG-binding site in the CH1 domain of mouse IgG(1) had been addressed. Interestingly, biosensor-based binding experiments showed that neither human IgG(1) nor bovine Ig, the latter present in fetal bovine serum (FBS) was recognized by Z(mab25). This selective binding profile towards murine IgG(1) was successfully exploited in species selective recovery of two different mouse mAbs from complex samples containing FBS, resembling a hybridoma culture supernatant.
Place, publisher, year, edition, pages
2011. Vol. 48, no 3, 263-276 p.
Mouse IgG1, Affibody, Selection, Fab fragment, Combinatorial protein engineering, Ribosome display
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-33435DOI: 10.1007/s12033-010-9367-1ISI: 000291656700008PubMedID: 21197589ScopusID: 2-s2.0-80054079725OAI: oai:DiVA.org:kth-33435DiVA: diva2:415482
FunderSwedish Research Council, 50548301
QC 201107052011-05-062011-05-062015-04-16Bibliographically approved