title: Cryoprotection of antibody by organic solutes and organic solute/divalent cation mixtures.
abstract: Antibodies are globular glycoproteins that protect animals from microbial and toxic insult. These proteins have proven to have substantial commercial and research value but are variably susceptible to freeze-thaw damage, thus limiting their usefulness. Several carbohydrates and divalent cations were examined alone and in combination to determine whether they could protect antibody from freeze-thaw damage. The amino acid proline was also tested. Two antibodies, derived from different sources and specific for different antigens, were tested by a direct enzyme-linked immunosorbent assay (ELISA). Confirmation of antibody freeze-lability was obtained by rocket electrophoresis and radial immunodiffusion tests. Neither carbohydrate nor divalent cation alone fully protected antibody activity from freeze-thaw damage. However, several combinations protected antibody activity completely when compared to their effect on antibody maintained at room temperature. In the case of affinity-purified antibody, full protection of antibody activity relative to an untreated control was obtained. In several instances, cryoprotection of antibody by solute-divalent cation combinations was synergistic and not an additive effect of each component. Alkaline phosphatase, an enzyme typically linked to antibody for an ELISA, was not freeze-thaw labile. These results indicate that antibody function can be fully protected from damage due to freeze-thaw treatment by organic solutes in combination with divalent cations.
authors.affiliation: Department of Microbiology, University of Southwestern Louisiana, Lafayette 70504-1007.
authors.forename: K C
authors.initials: KC
authors.lastname: Hazen
authors.forename: L D
authors.initials: LD
authors.lastname: Bourgeois
authors.forename: J F
authors.initials: JF
authors.lastname: Carpenter
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000906:Antibodies; D000937:Antigen-Antibody Reactions; D001666:Binding Sites, Antibody; D003451:Cryoprotective Agents; D005615:Freezing; D007120:Immunochemistry; D012997:Solvents; D014199:Trehalose; D015032:Zinc
chemical_list: D000906:Antibodies; D003451:Cryoprotective Agents; D012997:Solvents; D014199:Trehalose; D000469:Alkaline Phosphatase; D015032:Zinc