An improved method to create nitrocellulose particles suitable for the immobilization of antigen and antibody.
A method is described for producing 1-3 microns sized particles of nitrocellulose (NC) which are able to absorb protein. Protein is absorbed onto preformed particles made by first dissolving a sheet of nitrocellulose paper in DMSO, and then precipitating it with sodium carbonate/bicarbonate buffer. The efficiency of binding is the same as that of an equivalent sheet of non-processed NC filter paper. Antibodies absorbed onto preformed particles are not exposed to DMSO and carbonate buffer and therefore retain a high antigen binding capacity. Antigen and antibody-absorbed NC preformed particles were used to capture antibody and antigen, respectively. Using lysis buffer, the captured antibodies and antigens were readily released from the NC particles. This makes the latter an appropriate matrix for immunoprecipitation assays either for an antigen or for specific antibody. Antigen-coated NC particles were specifically aggregated ('agglutinated') by specific antibodies and thus can be used in semi-quantitative tests.
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D000936:Antigen-Antibody Complex / Q000276:immunology; D000941:Antigens / Q000302:isolation & purification*; D003101:Collodion / Q000737:chemistry*; D006801:Humans; D008863:Microspheres; D011233:Precipitin Tests / Q000379:methods*; D011485:Protein Binding; D011817:Rabbits
D000906:Antibodies; D000936:Antigen-Antibody Complex; D000941:Antigens; D003101:Collodion