Polyethylene Glycol (PEG)-Biotin antibody RabMAb®

Cat.#: 2137-1
Abcam ID: ab53449

Rabbit Monoclonal Antibody

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Description

Polyethylene glycol（PEG）is a family of long chain polymers attached to a glycerine backbone. It is a nonionic, nontoxic, biocompatible, strongly hydrophilic polymer, which has a large exclusion volume in aqueous solution (1). The covalent attachment of PEG is now commonly used to modify a variety of proteins and drugs (2,3). The modification of a biopharmaceutical with polyethylene glycol (PEG) increases its hydrodynamic radius, reduces immunogenicity and proteolytic cleavage. Other benefits include decelerated renal excretion, improved stability towards proteolysis and increased solubility of the biopharmaceutical in aqueous solutions (4). As examples, PEG-adenosine deaminase (Adagen) is used for the treatment of severe combined immunodeficiency syndrome, PEGasparaginase (Oncaspar) is used for the treatment acute lymphoblastic leukemia, PEGinterferon α2a (Pegasys) is used for the treatment Hepatitis C, Branched PEGanti-VEGF aptamer (Pegaptanib, Macugen) is used for the treatment Macular degeneration(age-related) (5). An Anti-PEG antibody can be used to monitor a drugs pharmacokinetics, including distribution, metabolism and excretion. In addition, it can be used for the quality control of pegylated molecules in ELISA, WB and flow cytometery.

Recommended Applications

WB, IHC, ELISA

Applications and Recommended Dilution Factors

WB: 15.6 ng/mL
IHC: 1:100
ELISA: 0.5 ug/ml

Species Reactivity *

*Cross reactivity determined by western blot only.

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Products Data

A. ELISA assay using 5ug/mL Anti-AFP RabMAb (#1791-1) as the capture antibody and 0.5 ug/mL Anti-PEG-47-Biotin RabMAb (cat. # 2137-1) as the detection antibody. HRP-avidin (1:3000) used for anti-PEG-47 detection. This ELISA assay could detect YCA1017 peptide-PEG antigen at a concentration as low as 0.9 ng/mL.	B. Sandwich ELISA assay using 5ug/mL Anti-GCSF as the capture antibody and 0.5 ug/mL Anti-PEG-47-Biotin RabMAb (cat. # 2137-1) as the detection antibody. HRP-avidin (1:3000) used for anti-PEG-47 detection. This ELISA assay could detect GCSF-PEG antigen at a concentration as low as 0.9 ng/mL.	C. Direct ELISA assay using 63 ng/mL of antigen (GCSF-PEG) to coat the plate. HRP-avidin (1:3000) was used for the detection of varying amounts of Anit-PEG-47-Biotin (cat. # 2137-1).
D. Immunoblot of different antigen-PEG preparations. Western Blot images using Anti-PEG 47-Biotin (cat. # 2137-1) to detect BSA-PEG. Lane 1: 31.25 ng/mL; lane 2: 15.6 ng/mL; lane 3: 7.8 ng/mL.

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Specificity

KLH-PEG with terminal methoxy group was used as an immunogen. This antibody recognizes the terminal methoxy group of the PEG molecule. Anti-PEG-47-Biotin is prepared by the biotinylation of the antibody via primary amines.

Storage Condition and Buffer

Store at -20 °C. Buffer: 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol, 0.01% sodium azide and 0.05% BSA. Stable for 12 months from date of receipt.

Description References

1. Guiotto, A. et al. (2004) Anchimeric assistance effect on regioselective hydrolysis of branched PEGs: a mechanistic investigation. Bioorg. Med.Chem. 12, 5031
2. Wong, S.S. (1991) Reactive groups of proteins and their modifying agents. In Chemistry of protein conjugation and cross-linking, p. 13, CRC Press
3. Caliceti, P. et al.(1993) Active site protection of proteolytic enzymes by poly(ethylene glycol) surface modification. J. Bioact. Comp. Polym. 8,41
4. Frank Leenders, celares GmbH, Berlin, Germany.(2006) PEGylation technology and biopharmaceuticals. Biopharmaceuticals. 6,39-40
5. Francesco M.Veronese, Gianfranco Pasut. (2005) PEGylation, successful approach