Baran M M, Allen D M, Russell S R, Scheetz M E, Monthony J F
J Immunol Methods. 1982 Sep 30;53(3):321-34. doi: 10.1016/0022-1759(82)90179-x.
Procedures are described for fractionating cells utilizing a universally applicable cellular affinity chromatography matrix. The affinity matrix consists of immunoabsorption purified goat anti-fluorescein isothiocyanate antibody coupled to large derivatized polyacrylamide beads. This matrix may, in principle, be used to isolate any cell subpopulation provided it has a fluorescein-labeled ligand on its surface. In this report the matrix was used to isolate viable purified fractions of mouse surface Ig-positive cells, Lyt1 cells, and mouse lymphocytes that bind the lectin soybean agglutinin. A preliminary experiment using the anti-FITC beads suggested that this technique can provide a fraction of cells enriched in antigen binding cells. Cell populations isolated by this technique retain their ability to respond to in vitro mitogen stimulation, as well as their ability to be maintained in cell culture following fractionation. Additional experiments using a column consisting of goat anti-rabbit Ig antibody coupled to the same support material are also reported.
描述了利用一种普遍适用的细胞亲和层析基质对细胞进行分级分离的方法。该亲和基质由免疫吸附纯化的山羊抗异硫氰酸荧光素抗体偶联到大型衍生化聚丙烯酰胺珠上组成。原则上，只要任何细胞亚群表面有荧光素标记的配体，这种基质就可用于分离该细胞亚群。在本报告中，该基质用于分离小鼠表面Ig阳性细胞、Lyt1细胞以及结合凝集素大豆凝集素的小鼠淋巴细胞的活纯化级分。使用抗FITC珠进行的初步实验表明该技术能够提供富含抗原结合细胞的细胞级分。通过该技术分离得到的细胞群体保留了对体外有丝分裂原刺激作出反应的能力，以及在分级分离后在细胞培养中维持的能力。还报告了使用偶联到相同支持材料上的山羊抗兔Ig抗体组成的柱进行的其他实验。
