Abstract
Phage display technology is frequently used to obtain antigen specific binders with predetermined characteristics. Phage display libraries are often constructed from animals immunized with the antigen of interest. An important point of consideration when making immune libraries is the availability of an appropriate antigen sources. When available, often either the amount is not sufficient for immunization or it is expensive to obtain. To overcome this problem, these antigens are typically obtained by over expression in prokaryotic or eukaryotic expression systems. While this could solve the problem of obtaining sufficient quantities of antigen for a reasonable price and effort, correct folding and differences in posttranslational modification could potentially lead to binders that recognize the recombinant, but not the endogenous protein. In addition, selection of binders against specific modifications or structural epitopes could be missed.
In this chapter we describe a particular selection of VHH antibody fragments from phage display libraries that were constructed from llamas immunized with different complex protein samples containing the antigen of interest. We show that this can result in binders that preferentially recognize the target of interest when present in specific structures depending on the antigen source.
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References
Marks JD, Hoogenboom HR, Bonnert TP et al (1991) By-passing immunization. Human antibodies from V-gene libraries displayed on phage. J Mol Biol 222:581–597
Roovers RC, van der Linden E, Zijlema H et al (2001) Evidence for a bias toward intracellular antigens in the local humoral anti-tumor immune response of a colorectal cancer patient revealed by phage display. Int J Cancer 93:832–840
Sommavilla R, Lovato V, Villa A et al (2010) Design and construction of a naïve mouse antibody phage display library. J Immunol Methods 353:31–43
Chiliza TE, Van Wyngaardt W, Du Plessis DH (2008) Single-chain antibody fragments from a display library derived from chickens immunized with a mixture of parasite and viral antigens. Hybridoma 27:413–421
van Koningsbruggen S, de Haard H, de Kievit P et al (2003) Llama-derived phage display antibodies in the dissection of the human disease oculopharyngeal muscular dystrophy. J Immunol Methods 279:149–161
Fu Y, Shearing LN, Haynes S et al (1997) Isolation from phage display libraries of single chain variable fragment antibodies that recognize conformational epitopes in the malaria vaccine candidate, apical membrane antigen-1. J Biol Chem 272:25678–25684
Sahdev S, Khattar SK, Saini KS (2008) Production of active eukaryotic proteins through bacterial expression systems: a review of the existing biotechnology strategies. Mol Cell Biochem 307:249–264
Lang IM, Barbas CF III, Schleef RR (1996) Recombinant rabbit Fab with binding activity to type-1 plasminogen activator inhibitor derived from a phage-display library against human alpha-granules. Gene 172:295–298
Roovers RC, Laeremans T, Huang L et al (2007) Efficient inhibition of EGFR signaling and of tumour growth by antagonistic anti-EFGR nanobodies. Cancer Immunol Immunother 56:303–317
Omidfar K, Rasaee MJ, Modjtahedi H et al (2004) Production of a novel camel single-domain antibody specific for the type III mutant EGFR. Tumour Biol 25:296–305
Rutgers KS, van Remoortere A, van Buchem MA et al (2011) Differential recognition of vascular and parenchymal beta amyloid deposition. Neurobiol Aging 32:1774–1783
Bornebroek M, Van Buchem MA, Haan J et al (1996) Hereditary cerebral hemorrhage with amyloidosis-Dutch type: better correlation of cognitive deterioration with advancing age than with number of focal lesions or white matter hyperintensities. Alzheimer Dis Assoc Disord 10:224–231
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, vol 1–3. Cold Spring Harbor Laboratory Press, New York
Acknowledgements
This work was supported by grants from the IOP Genomics Center (IGE05005), the National Institutes of Health (NIH AG021084 and AG005134) and by the Centre for Medical Systems Biology within the framework of the Netherlands Genomics Initiative (NGI)/Netherlands Organization for Scientific Research (NWO).
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Klooster, R., Rutgers, K.S., van der Maarel, S.M. (2012). Selection of VHH Antibody Fragments That Recognize Different Aβ Depositions Using Complex Immune Libraries. In: Saerens, D., Muyldermans, S. (eds) Single Domain Antibodies. Methods in Molecular Biology, vol 911. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-968-6_15
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DOI: https://doi.org/10.1007/978-1-61779-968-6_15
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