Covalent linking of haptens, proteins and nucleic acids to a modified polystyrene support

• Published in: Journal of immunological methods Vol. 159; no. 1; pp. 177 - 187
• Main Authors: Niveleau, A., Sage, D., Reynaud, C., Bruno, C., Legastelois, S., Thomas, V., Dante, R.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 26.02.1993; Elsevier
• Subjects: Biological and medical sciences; Covalent; ELISA; Enzyme-Linked Immunosorbent Assay - methods; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Hapten; Haptens - analysis; Molecular immunology; Nucleic Acids - analysis; Polynucleotide; Polystyrene; Polystyrenes; Protein; Proteins - analysis; Techniques; PBS; AMPPD; poly C; Polynucleotide; EDC; Bio-4-dUTP; Polystyrene; Hapten; Protein; Covalent; NHS-biotin; PBST; NHS; poly I; MES; PNPP; TCT buffer; DMF; ELISA; Proteins; Immobilization; Covalent bond; Styrene polymer; Solid phase; ELISA assay; Nucleic acid
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/0022-1759(93)90156-2

Biological molecules of various molecular weights were successfully linked to polystrrene microtitration plates bearing carbonyl and hydroxyl reactive groups. Cross-linking of proteins, nucleic acids and haptens was carried out with water soluble carbodiimide. The covalent nature of the binding reaction was demonstrated by the immunodetection of a hapten linked to the surface through a disulphide spacer arm and its subsequent release by cleavage of the bridge. The amount of protein fixed per surface unit could be correlated to molecular weight. Nanograms of biotinylated nucleic acids and synthetic polynucleotides could also be retained on the solid support.