Evaluation of apparent non-specific protein loss due to adsorption on sample tube surfaces and/or altered immunogenicity.
The non-specific loss of protein analytes can have a major effect on assay results particularly where the concentrations of such analytes are extremely low and the matrix is complex. This report assesses how the protein incubated in sample tubes may be lost due to adsorption. Use of proteins, such as bovine serum albumin (BSA), may be used to pre-treat tubes to reduce such losses. However, such losses may also be associated with structural perturbations leading to changes in immunogenicity (as a result of alterations in specific epitope-related conformations). This can lead to erroneous results or lack of comparability with a range of methodologies such as the bicinchoninic protein assay and immunoassays or when surface plasmon resonance (SPR)-based approaches are used. A model system to evaluate these phenomena is proposed.
Centre for Bioanalytical Sciences (CBAS), National Centre for Sensor Research, Dublin City University, Dublin 9, Ireland.
Chandra K
CK
Dixit
Sandeep K
SK
Vashist
Brian D
BD
MacCraith
Richard
R
O'Kennedy
D000327:Adsorption; D000818:Animals; D001798:Blood Proteins / Q000737:chemistry / Q000276:immunology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005898:Glass / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D051379:Mice; D011126:Polypropylenes / Q000737:chemistry; D011804:Quinolines / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D020349:Surface Plasmon Resonance / Q000379:methods*; D013499:Surface Properties; D060749:alpha-2-HS-Glycoprotein
C556354:AHSG protein, human; C556357:Ahsg protein, mouse; D001798:Blood Proteins; D007074:Immunoglobulin G; D011126:Polypropylenes; D011804:Quinolines; D060749:alpha-2-HS-Glycoprotein; D012710:Serum Albumin, Bovine; C047117:bicinchoninic acid; D006735:Horseradish Peroxidase