SpringerProtocols: Abstract: Rationally Designed Ligands for Use in Affinity Chromatography: An Artificial Protein L

7. Rationally Designed Ligands for Use in Affinity Chromatography:
An Artificial Protein L

By: Ana Cecília A. Roque², Christopher R. Lowe³

Abstract

Full Text

Download PDF (550K)

Synthetic affinity ligands can circumvent the drawbacks of natural immunoglobulin (Ig)-binding proteins by imparting resistance to chemical and biochemical degradation and to in situ sterilization, as well as ease and low cost of production. Protein L (PpL), isolated from Peptostreptococcus magnus strains, interacts with the Fab (antigen-binding fragment) portion of Igs, specifically with kappa light chains, and represents an almost universal ligand for the purification of antibodies. The concepts of rational design and solid-phase combinatorial chemistry were used for the discovery of a synthetic PpL mimic affinity ligand. The procedure presented in this chapter represents a general approach with the potential to be applied to different systems and target proteins.

Affiliation(s): (2) Faculdade de Ciéncias e Tecnologia, Universidade Nova de Lisboa, Portugal
(3) Department of Biotechnology, Institute of Biotechnology, University of Cambridge, Cambridge, UK

Book Title: Affinity Chromatography: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 421 | Pub. Date: Nov-30-2007 | Page Range: 93-110 | DOI: 10.1007/978-1-59745-582-4_7

Subject: Cell Biology

Key Words: Affinity - biomimetic - ligands - synthetic - proteins - purification - design - combinatorial synthesis - screening - Protein L

References

Download References

1.	Lowe, C. R., Lowe, A. R. and gupta, g. (2001) New developments in affinity chromatography with potential application in the production of biopharmaceuticals. J. Biochem. Biophys. Methods 49, 561–574.

2.	Stockwin, L. and Holmes, S. (2003) Antibodies as therapeutic agents: vive la renaissance! Expert Opin. Biol. Ther. 3, 1133–1152.

3.	Huse, K., Bohme, H. J. and Scholz, g. H. (2002) Purification of antibodies by affinity chromatography. J. Biochem. Biophys. Methods 51, 217–231.

4.	Roque, A. C. A., Lowe, C. R. and Taipa, M. A. (2004) Antibodies and genetically engineered related molecules: production and purification. Biotechnol. Prog. 20, 639–654.

5.	Fassina, g., Verdoliva, A., Odierna, M. R., Ruvo, M. and Cassini, g. (1996) Protein a mimetic peptide ligand for affinity purification of antibodies. J. Mol. Recognit. 9, 564–569.

6.	Li, R. X., Dowd, V., Stewart, D. J., Burton, S. J. and Lowe, C. R. (1998) Design, synthesis, and application of a Protein A mimetic. Nat. Biotechnol. 16, 190–195.

7.	Housden, N. g., Harrison, S., Roberts, S. E., Beckingham, J. A., graille, M., Stura, E. A. and gore, M. g. (2003) Immunoglobulin-binding domains: protein L from Peptostreptococcus magnus. Biochem. Soc. Trans. 31, 716–718.

8.	Stura, E. A., graille, M., Housden, N. g. and gore, M. g. (2002) Protein L mutants for the crystallization of antibody fragments. Acta Crystallogr. Sect. D Biol. Crystallogr. 58, 1744–1748.

9.	Lowe, C. R., Burton, S. J., Burton, N. P., Alderton, W. K., Pitts, J. M. and Thomas, J. A. (1992) Designer dyes - biomimetic ligands for the purification of pharmaceutical proteins by affinity-chromatography. Trends Biotechnol. 10, 442–448.

10.	Graille, M., Stura, E. A., Housden, N. g., Beckingham, J. A., Bottomley, S. P., Beale, D., Taussig, M. J., Sutton, B. J., gore, M. g. and Charbonnier, J. B. (2001) Complex between Peptostreptococcus magnus protein L and a human antibody reveals structural convergence in the interaction modes of Fab binding proteins. Structure 9, 679–687.

11.	Boeijen, A. and Liskamp, R. M. J. (1999) Solid-phase synthesis of oligourea peptidomimetics. Eur. J. Org. Chem. 2127–2135.

12.	Nilson, B. H. K., Logdberg, L., Kastern, W., Bjorck, L. and Akerstrom, B. (1993) Purification of antibodies using Protein-L-binding framework structures in the light-chain variable domain. J. Immunol. Methods 164, 33–40.

13.	Roque, A. C. A., Taipa, M. A. and Lowe, C. R. (2004) A new method for the screening of solid-phase combinatorial libraries for affinity chromatography. J. Mol. Recognit. 17, 262–267.

14.	Roque, A. C. A., Taipa, M. A. and Lowe, C. R. (2005) Synthesis and screening of a rationally designed combinatorial library of affinity ligands mimicking protein L from Peptostreptococcus magnus. J. Mol. Recognit. 18, 213–224.

15.	Roque, A. C. A., Taipa, M. A. and Lowe, C. R. (2005) An artificial protein L for the purification of immunoglobulins and Fab fragments by affinity chromatography. J. Chromatogr. A 1064, 157–167.

16.	Snyder, S. L. and Sobocinski, P. Z. (1975) An improved 2,4,6-trinitro benzenesulfonic acid method for the determination of amines. Anal. Biochem. 64, 284–288.

17.	Kaiser, E., Colescott, R., Bossinger, C. D. and Cook, P. I. (1970) Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides. Anal. Biochem. 34, 595–598.

18.	Dam, T. K., Roy, R., Page, D. and Brewer, C. F. (2002) Negative cooperativity associated with binding of multivalent carbohydrates to lectins. Thermodynamic analysis of the “multivalency effect”. Biochemistry 41, 1351–1358.

19.	Ohno, K., Fukushima, T., Santa, T., Waizumi, N., Tokuyama, H., Maeda, M. and Imai, K. (2002) Estrogen receptor binding assay method for endocrine disruptors using fluorescence polarization. Anal. Chem. 74, 4391–4396.

20.	Munson, P. J. and Rodbard, D. (1980) LIGAND: a versatile computerized approach for characterization of ligand-binding systems. Anal. Biochem. 107, 220–239.

Comments (Loading...)

Post comment/View all comments