Monoclonal antibody purification by affinity chromatography with ligands derived from the screening of peptide combinatory libraries

Biotechnology Letters - Tập 25 - Trang 1545-1548 - 2003
S.A. Camperi1, N.B. Iannucci1, G.J. Albanesi1, M. Oggero Eberhardt2, M. Etcheverrigaray2, A. Messeguer3, F. Albericio4, O. Cascone1
1Cátedra de Microbiología Industrial y Biotecnología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina
2Laboratorio de Cultivos Celulares, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, Argentina
3Instituto de Investigaciones Bioquímicas y Ambientales (CSIC), Barcelona, Spain
4Departamento de Química Orgánica, Facultad de Química, Universidad de Barcelona, Barcelona, Spain

Tóm tắt

The peptide, Ala-Pro-Ala-Arg (APAR), was selected from the screening of a tetrapeptide combinatorial synthetic library as the ligand for affinity purification of an anti-Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF) monoclonal antibody (Mab) developed in mouse ascitis. The affinity chromatographic matrix obtained by attachment of APAR to agarose, having a peptide density of 0.5 μmol ml−1, showed a maximum capacity of 9.1 mg Mab ml−1 and a dynamic capacity of 3.9 mg Mab ml−1. A 95% yield of electrophoretically pure anti-GM-CSF was obtained in a single step.

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Tài liệu tham khảo

Baumbach GA, Hammond DJ (1993) Protein purification using af-finity ligands deduced from peptide libraries. J. Cell. Biochem. Suppl 17A: 43.

Chase HA (1984) Prediction of the performance of preparative affinity chromatography. J. Chromatog. 297: 179–202.

Cwirla SE, Peters EA, Barrett RW, Dower WJ (1990) Peptides on phage: a vast library of peptides for identifying ligands. Proc. Natl. Acad. Sci. USA 87: 6378–6382.

Hermanson GT, Mallia KA, Smith PK (1992) Immobilization of ligands. In: Immobilized Affinity Ligand Techniques. San Diego, USA: Academic Press.

Hirabayashi Y, Fukuda H, Kimura J, Miyamoto M, Yasui K (1996) Identification of peptides mimicking the antigenicity and immunogenicity of conformational epitopes on Japanese encephalitis virus protein using synthetic peptide libraries. J. Virol. Meth. 61: 23–36.

Huang PY, Carbonell RG (1999) Affinity chromatographic screening of soluble combinatorial peptide libraries. Biotechnol. Bioeng. 63: 633–641.

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 22: 680–685.

Lam KS, Salmon SE, Hersh EM, Hruby VJ, Kazmierski WM(1991) A new type of synthetic peptide library for identifying ligandbinding activity. Nature 354: 82–84.

Lloyd-Williams P, Albericio F, Giralt E (1997) In: Chemical Approaches to the Synthesis of Peptides and Proteins. Boca Raton, FL: CRC Press.

Lowe CR, Pearson JC (1984) Affinity chromatography on immobilized dyes. Meth. Enzymol. 104: 97–113.

Oggero Eberhardt M, Frank R, Kratje R, Etcheverrigaray M (2003) Identification of two potential receptor-binding sites for hGM- CSF. Brazilian J. Chem. Eng. 20: 15–20.

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Biotechnology Letters

Tập 25

1545-1548

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