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INSTITUTO DE PESQUISAS ENERGÉTICAS E NUCLEARES Autarquia associada à Universidade de São Paulo ESTUDO DA MARCAÇÃO E BIODISTRIBUIÇÃO DA SUBSTÂNCIA P UTILIZANDO LUTÉCIO-177 COMO RADIOTRAÇADOR
| Content Provider | Semantic Scholar |
|---|---|
| Author | Lima, Clarice Maria De |
| Copyright Year | 2011 |
| Abstract | Purification of human prolactin from pituitaries was carried out in our laboratory to obtain a pure reagent for use in RIA. The extraction and purification procedure was adapted from the method of Mc. Lean et al., and it involves the following steps: 1= Extraction of frozen pituitaries in buffers 0.i4i^ phosphate/citrate pH 4.0 and 0.05M ammonium acetate pH i0.0. 2. Purification by hydrophobic interaction chromatography on Pheny l-Sepharose CL-4i3 in the presence of aceton i tr i le. 3. Purification by anion exchange chromatography on DEAESepharosc CL-6B. The whole process from the first homogenization step to the starting of the 1yophi1ization can be performed in about 10 days. In the extraction phase an extra step was added using buffer 50mi^ ammonium acetate pH 7.0, which allowed a significant enrichment in the hPrl content of the extract. Prolactin was recovered from 30-70 pituitaries with an overall yield of approximately 7 /ig/p i tu i t ar ie. The purity of the hormone was analyzed on 7% polyacrilamide gel electrophoresis. hPrl-IPEN presented a single band with Rm=0.505, practically coincident with the Rm=0.503 of NIADDKhPrl-I-7. In the hPrl RIA, the standard curves obtained with hPrl-IPEN and NIADDK-hPr1-1-7 showed a significant parallelism (P<0.05), By specific RIA, the purest hPrl-IPEN presented only 0.7% of immunoreactive hGH. A work was carried out also on internal quality control of hPrl-RIA. First, using hPrl from NIADDK, a study was performed on radioiodination reprodutibi1ity, as well as sensitivity, precision and accuracy of the assays. Then, once obtained the hPrl-IPEN standard preparation, the same type of quality control was carried out on this ampolized reference material. The hPrl immunoreactive content was of 492.0 ug/ampoule (CV inter assay, inter ampoule=i3.2%), its stability was controlled for five months, not showing any significant decrease, while the precision of the assay was of the same order of that obtained with hPrl NIADDK. The purification method is considered effective for obtaining a hPrl of the purity needed for radioassay purposes, having the advantage of rapidity and relative simplicity. |
| File Format | PDF HTM / HTML |
| DOI | 10.11606/D.85.2011.tde-05082011-102555 |
| Alternate Webpage(s) | http://pelicano.ipen.br/PosG30/TextoCompleto/Ligia%20Ely%20Morganti%20Ferreira%20Dias_M.pdf |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
