دورية أكاديمية
Anti-EPO and anti-NESP antibodies raised against synthetic peptides that reproduce the minimal amino acid sequence differences between EPO and NESP.
| العنوان: | Anti-EPO and anti-NESP antibodies raised against synthetic peptides that reproduce the minimal amino acid sequence differences between EPO and NESP. |
|---|---|
| المؤلفون: | Giménez E; Departament de Química Analítica, Universitat de Barcelona, Diagonal 647, 08028 Barcelona, Spain., de Bolós C, Belalcazar V, Andreu D, Borrás E, De la Torre BG, Barbosa J, Segura J, Pascual JA |
| المصدر: | Analytical and bioanalytical chemistry [Anal Bioanal Chem] 2007 Aug; Vol. 388 (7), pp. 1531-8. Date of Electronic Publication: 2007 May 30. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Springer-Verlag Country of Publication: Germany NLM ID: 101134327 Publication Model: Print-Electronic Cited Medium: Print ISSN: 1618-2642 (Print) Linking ISSN: 16182642 NLM ISO Abbreviation: Anal Bioanal Chem Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Heidelberg : Springer-Verlag, 2002- |
| مواضيع طبية MeSH: | Antibodies, Monoclonal*/biosynthesis, Erythropoietin/*immunology , Peptide Fragments/*immunology, Amino Acid Sequence ; Darbepoetin alfa ; Epitopes ; Erythropoietin/analogs & derivatives ; Erythropoietin/analysis ; Humans ; Immunoassay ; Peptide Fragments/chemical synthesis ; Recombinant Proteins |
| مستخلص: | Erythropoietin (EPO) is a hormone that regulates red blood cell production. Recombinant human EPO (rHuEPO) and NESP (novel erythropoiesis stimulating protein) have been produced for therapeutic purposes and also to improve sports performance. The primary sequences of rHuEPO and NESP differ by just five amino acids. Due to the high homology, no antibodies that are able to discriminate between both molecules have been obtained until now. The aim of the present work was to design synthetic peptides corresponding to the sequence that differs between EPO and NESP (87-90aa), that can then be used as immunogens to develop specific rabbit polyclonal antibodies for selectively detecting EPO and NESP. Three peptides were synthesized: EPO (81-95), NESP (81-95), and NESP (86-104), and these were coupled to KLH and OVA for immunization and screening purposes, respectively. The sera obtained were tested by ELISA on synthetic peptide-OVA conjugates and purified by immunoaffinity chromatography against the corresponding synthetic peptide. The specific purified antibodies were characterized by ELISA, SDS-PAGE, and isoelectric focusing, followed by western blot. Antisera raised against EPO (81-95) recognized rHuEPO but not NESP. In contrast, anti-NESP (84-106) sera gave a specific anti-NESP response only after immunoaffinity purification on a NESP (86-91) column. An efficient strategy for generating specific antibodies against EPO and NESP can be achieved by selecting suitable synthetic peptides. The antibodies obtained are able to differentiate between rHuEPO and NESP, and may be particularly useful for screening purposes in both therapeutic and antidoping contexts. |
| المشرفين على المادة: | 0 (Antibodies, Monoclonal) 0 (Epitopes) 0 (Peptide Fragments) 0 (Recombinant Proteins) 11096-26-7 (Erythropoietin) 15UQ94PT4P (Darbepoetin alfa) |
| تواريخ الأحداث: | Date Created: 20070531 Date Completed: 20070905 Latest Revision: 20160512 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1007/s00216-007-1334-8 |
| PMID: | 17534608 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Full Text Finder كن أول من يترك تعليقا!