Adamus G, Zam Z S, Emerson S S, Hargrave P A
Department of Ophthalmology, University of Florida, Gainesville 32610.
In Vitro Cell Dev Biol. 1989 Dec;25(12):1141-6. doi: 10.1007/BF02621265.
We present a practical method for the rescue of previously stable hybridoma clones which increases the proportion of desired cells in the population before cloning by limiting dilution. When the antibody activity of a culture supernatant was lower than that previously obtained, a precloning distribution at a density of 10 cells per microtiter well greatly improved the chances of obtaining a single active clone by subsequent limiting dilution. The Poisson distribution model was used to evaluate the method. Probabilities calculated clearly demonstrate the advantage of this precloning distribution step when attempting to isolate a hybridoma cell line that is relatively rare in a population.
我们提出了一种实用方法，用于挽救先前稳定的杂交瘤克隆，该方法通过有限稀释在克隆前增加群体中所需细胞的比例。当培养上清液的抗体活性低于先前获得的活性时，以每微滴孔10个细胞的密度进行预克隆分布，可大大提高通过后续有限稀释获得单个活性克隆的几率。泊松分布模型用于评估该方法。计算出的概率清楚地证明了在试图从群体中分离相对稀有的杂交瘤细胞系时，这一预克隆分布步骤的优势。
