Adsorption of EGF receptor ligands to test tubes--a factor with implications for studies on the potency of these peptides.
Studies on the relative potency of ligands for the epidermal growth factor receptor are usually performed with highly purified ligand specimens. However, adsorption of ligands to glass and plastic surfaces may affect the results by reducing the ligand concentration in an unpredictable way. The aim of this study was to examine the adsorption of four epidermal growth factor (EGF) receptor ligands, EGF, transforming growth factor alpha (TGF-alpha), heparin binding-EGF (HB-EGF) and betacellulin, to commonly used test tubes of polyethylene, polystyrene and glass, respectively. The ligands were kept in a sodium phosphate buffer, both with and without 0.1% human albumin as carrier protein. Adsorption was examined after 20 minutes at room temperature as well as after overnight storage at 4 degrees C. The ligands were quantitated by ELISAs. In the buffer not containing 0.1% human albumin there was a marked adsorption, which differed both among the ligands and among the test tubes. After 20 minutes the ligand concentrations were reduced to 33-73% in polyethylene tubes, to 15-46% in polystyrene tubes and to 12-29% in glass tubes. The adsorption was even more pronounced after storage overnight. The use of 0.1% human albumin in the buffer solved the problem in polyethylene and polystyrene tubes, but not in glass tubes. The results demonstrate that adsorption to surfaces can be a significant problem for EGF receptor ligands and emphasize the need for controlling the growth factor concentration in the final experimental setting.
Department of Clinical Biochemistry, AKH Aarhus University Hospital, Denmark.
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Jørgensen
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Eskildsen
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Nexø
D000327:Adsorption; D066259:Betacellulin; D002624:Chemistry, Clinical / Q000592:standards; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards; D004815:Epidermal Growth Factor / Q000032:analysis / Q000493:pharmacokinetics* / Q000494:pharmacology; D066246:ErbB Receptors / Q000378:metabolism*; D005898:Glass; D006133:Growth Substances / Q000032:analysis / Q000493:pharmacokinetics / Q000494:pharmacology; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D011095:Polyethylenes; D011137:Polystyrenes; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D015203:Reproducibility of Results; D016211:Transforming Growth Factor alpha / Q000032:analysis / Q000493:pharmacokinetics / Q000494:pharmacology
C587851:BTC protein, human; D066259:Betacellulin; D006133:Growth Substances; C587883:HBEGF protein, human; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D011095:Polyethylenes; D011137:Polystyrenes; D011994:Recombinant Proteins; D016211:Transforming Growth Factor alpha; D004815:Epidermal Growth Factor; D066246:ErbB Receptors