Purificación de cinco isoformas de la hormona luteinizante bovina (bLH). Caracterización fisicoquímica, biológica e inmunológica
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| Format: | Artículo científico |
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Universidad Nacional Autónoma de México
2004
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| author | Andrés Falcón Alcántara |
| author_facet | Andrés Falcón Alcántara |
| contents | Purificación de cinco isoformas de la hormona luteinizante bovina (bLH). Caracterización fisicoquímica, biológica e inmunológica Andrés Falcón Alcántara Joel Hernández Cerón Everardo González Padilla Clara Murcia Mejía Gerardo Perera Marín Multidisciplinaria (Ciencias Naturales y Exactas) bovine pituitary ionic interchange lutenizing hormone (lh) isoforms The study describes the purifi cation of fi ve LH isoforms from bovine pituitary extracts. Cationic interchangechromatography was used to obtain fractions at a pH of 6.8 and 9.5. These were then repurifi edunder identical conditions in DEAE-Sephacel to obtain LH-I (1.2 mg/kg of pituitary), and LH-II (1.8mg/kg), LH-III (1.5 mg/kg), LH-IV (1.2 mg/kg) and LH-V (7.0 mg/kg), respectively. The relative mobility(Rf) of each isoform was determined using polyacrylamide gel electrophoresis under native conditions.LH-I presented an Rf of 0.16 cm; LH- II, LH-III and LH-IV had an Rf of 0.044 cm, while LH-Vhad a 0.11 cm Rf. The molecular weight (MW) of LH-I and LH-V was 37.5 kDa, while LH-II, LH-III andLH-IV had a MW of 38.5 kDa. The immunotransference analysis identifi ed MW bands weighing 59.0,37.5 and 22.0 kDa in the standards as well as in the different isoforms. The in vivo biological potencycorresponded to 1.04 U/mg (LH-I); 5.9 U/mg (LH-II); 0.16 U/mg (LH-III); 0.86 U/mg (LH-IV) and 1.4U/mg (LH-V) and their LH radioinmmunoassay (RIA) 50% expected dose (ED50) were 2.27 ng/tube(LH-I), 1.19 ng/tube (LH-II), 1.35 ng/tube (LH-III), 0.66 ng/tube (LH-IV) and 1.71 ng/tube (LH-V).These data show that it is possible to obtain two LH isoforms at pH 6.8 (I and V) and three at pH 9.5(II, III and IV), with heterogeneous charges, RIA and biological activity, with suffi cient yield as to try todevelop simpler and more precise analytical methods. 2004 artículo científico 0301-5092 https://www.redalyc.org/articulo.oa?id=42335205 en http://www.redalyc.org/revista.oa?id=423 Veterinaria México application/pdf Universidad Nacional Autónoma de México Veterinaria México (México) Num.2 Vol.35 |
| format | Artículo científico |
| id | redalyc_42335205 |
| institution | Redalyc |
| language | en |
| publishDate | 2004 |
| publisher | Universidad Nacional Autónoma de México |
| spellingShingle | Purificación de cinco isoformas de la hormona luteinizante bovina (bLH). Caracterización fisicoquímica, biológica e inmunológica Andrés Falcón Alcántara Multidisciplinaria (Ciencias Naturales y Exactas) bovine pituitary ionic interchange lutenizing hormone (lh) isoforms Purificación de cinco isoformas de la hormona luteinizante bovina (bLH). Caracterización fisicoquímica, biológica e inmunológica Andrés Falcón Alcántara Joel Hernández Cerón Everardo González Padilla Clara Murcia Mejía Gerardo Perera Marín Multidisciplinaria (Ciencias Naturales y Exactas) bovine pituitary ionic interchange lutenizing hormone (lh) isoforms The study describes the purifi cation of fi ve LH isoforms from bovine pituitary extracts. Cationic interchangechromatography was used to obtain fractions at a pH of 6.8 and 9.5. These were then repurifi edunder identical conditions in DEAE-Sephacel to obtain LH-I (1.2 mg/kg of pituitary), and LH-II (1.8mg/kg), LH-III (1.5 mg/kg), LH-IV (1.2 mg/kg) and LH-V (7.0 mg/kg), respectively. The relative mobility(Rf) of each isoform was determined using polyacrylamide gel electrophoresis under native conditions.LH-I presented an Rf of 0.16 cm; LH- II, LH-III and LH-IV had an Rf of 0.044 cm, while LH-Vhad a 0.11 cm Rf. The molecular weight (MW) of LH-I and LH-V was 37.5 kDa, while LH-II, LH-III andLH-IV had a MW of 38.5 kDa. The immunotransference analysis identifi ed MW bands weighing 59.0,37.5 and 22.0 kDa in the standards as well as in the different isoforms. The in vivo biological potencycorresponded to 1.04 U/mg (LH-I); 5.9 U/mg (LH-II); 0.16 U/mg (LH-III); 0.86 U/mg (LH-IV) and 1.4U/mg (LH-V) and their LH radioinmmunoassay (RIA) 50% expected dose (ED50) were 2.27 ng/tube(LH-I), 1.19 ng/tube (LH-II), 1.35 ng/tube (LH-III), 0.66 ng/tube (LH-IV) and 1.71 ng/tube (LH-V).These data show that it is possible to obtain two LH isoforms at pH 6.8 (I and V) and three at pH 9.5(II, III and IV), with heterogeneous charges, RIA and biological activity, with suffi cient yield as to try todevelop simpler and more precise analytical methods. 2004 artículo científico 0301-5092 https://www.redalyc.org/articulo.oa?id=42335205 en http://www.redalyc.org/revista.oa?id=423 Veterinaria México application/pdf Universidad Nacional Autónoma de México Veterinaria México (México) Num.2 Vol.35 |
| title | Purificación de cinco isoformas de la hormona luteinizante bovina (bLH). Caracterización fisicoquímica, biológica e inmunológica |
| topic | Multidisciplinaria (Ciencias Naturales y Exactas) bovine pituitary ionic interchange lutenizing hormone (lh) isoforms |
| url | https://www.redalyc.org/articulo.oa?id=42335205 |