Sobsey M D, Lavigne M E, Cooper R C
Appl Microbiol. 1972 Mar;23(3):623-7. doi: 10.1128/am.23.3.623-627.1972.
Phosphorus-33 ((33)P), a weak (0.25 Mev) beta-emitting isotope of phosphorus with a half-life of 25 days, has been used to label poliovirus in cell culture. HeLa cell monolayers were depleted of phosphate and then labeled by incubating at 37 C in a medium (LM) containing about 10 muCi of (33)P as orthophosphate per ml. Labeled cells were infected at a high multiplicity with poliovirus type 1 and incubated for 8 hr in LM medium. Virus from infected cells was then concentrated and purified. Virus purity was confirmed by comparison of virus infectivity and radioactivity after CsCl density gradient centrifugation and by observing purified virus preparations with electron microscopy. With the method described, yields of about 10(10) to 5 x 10(10) plaque-forming units (PFU) of highly purified poliovirus with specific activities of about 3 x 10(-4) to 10(-3) disintegrations per min per PFU have been obtained from 1.5 x 10(8) to 3.0 x 10(8) HeLa cells.
磷 - 33（³³P）是磷的一种弱β发射同位素（0.25兆电子伏特），半衰期为25天，已被用于在细胞培养中标记脊髓灰质炎病毒。将HeLa细胞单层中的磷酸盐耗尽，然后在含有每毫升约10微居里正磷酸盐形式的³³P的培养基（LM）中于37℃孵育进行标记。用1型脊髓灰质炎病毒以高感染复数感染标记的细胞，并在LM培养基中孵育8小时。然后将来自感染细胞的病毒浓缩并纯化。通过在氯化铯密度梯度离心后比较病毒感染性和放射性以及用电镜观察纯化的病毒制剂来确认病毒纯度。用所述方法，从1.5×10⁸至3.0×10⁸个HeLa细胞中获得了产量约为10¹⁰至5×10¹⁰个空斑形成单位（PFU）的高度纯化的脊髓灰质炎病毒，其比活性约为每PFU每分钟3×10⁻⁴至10⁻³次衰变。
