Isolation of IgG immunoglobulins from rabbit alv-antiserum using protein-A-sepharose and estimation of globe artichoke local cultivar "istra" for presence of artichoke latent virus (alv) and cucumber mosaic virus (cmv) in Slovenia
1995
Baricevic, D. | Kus, M. | Pepelnjak, M. (Ljubljana Univ. (Slovenia). Biotechnical Fac., Agronomy Dept.)
Local globe artichoke cultivar 'Istra' was tested for presence of viruses ALV and CMV (serological type CuMV-cr and Cu Mv-vi) by means of biological testing (inoculation of indicator plants with sap of examined plant material) and by means of Enzyme - linked immunosorbent-assay (ELISA). The procedure of ALV virus determination included isolation of IgG fraction from rabbit antiserum using protein - A - Sepharose 4B column chromatography instead of standard IgG purification technique (DEAE Sephacel column chromatography). The principle of applied method lies in pH dependent regulation of eluation of gel-unbound or gel-bound proteins, either with phospate buffer (pH= 8.2) or citrate buffer (pH = 3.5) respectively. Protein - A -Sepharose 4B column chromatography is faster method of antiserum purification than DEAE Sephacel column chromatography so also ELISA testing is faster. Biological testing and ELISA procedure could't prove infection of examined plant material by ALV or CMV viruses
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