Expression of bovine inhibin β subunit in Escherichia coli

Kim, K.D.; Knight, P. G.; Savva, D.

doi:10.1016/0020-711x(91)90232-c

Cited by 3 publications

(1 citation statement)

References 33 publications

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“…The assay was calibrated using the same 32 kDa bovine inhibin standard that was used in the IRMA and total RIA. This assay has been used previously to confirm the expression of recombinant inhibin ß subumt protein by transformed E. coli containing the ß subunit gene (Kim et al 1991). …”

Section: Antibodiesmentioning

confidence: 99%

Measurement of dimeric inhibin using a modified two-site immunoradiometric assay specific for oxidized (Met O) inhibin

Knight

Muttukrishna²

1994

Journal of Endocrinology

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Several years ago we developed a novel two-site immunoradiometric assay (IRMA) for dimeric inhibin. However, relative to the purified 32 kDa bovine inhibin standard used at that time, the immunopotencies of crude inhibin-containing samples were much less than their biopotencies estimated by pituitary cell bioassay. In attempting to improve assay performance and resolve this discrepancy we recently discovered that introduction of a preassay oxidation step to the IRMA results in a dramatic increase in the immunopotencies of inhibin-containing test samples (e.g.: bovine, human, porcine follicular fluid (FF)) and of a new (purified in 1993) 32 kDa bovine inhibin standard. However, the oxidation step did not affect the immunopotency of our original standard (purified in 1987), indicating that this material had undergone spontaneous oxidation during long-term storage, thus accounting for its higher immunopotency in our original IRMA and providing an explanation for the discrepancy between immunoactivity and bioactivity referred to above. These findings, together with other observations on the behaviour of oxidized and non-oxidized samples of inhibin, related peptide fragments and inhibin-containing samples in the IRMA and alpha subunit radioimmunoassay (RIA), indicate that the anti-beta A82-114 monoclonal antibody (E4) used as tracer in the IRMA binds selectively to the oxidized (Met O89,91,108) form of the peptide. This property of the antibody can be exploited to advantage by incorporating simple modifications to existing inhibin/activin immunoassays to ensure that all samples and standards are fully oxidized before antibody addition.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Antibodiesmentioning

confidence: 99%

Measurement of dimeric inhibin using a modified two-site immunoradiometric assay specific for oxidized (Met O) inhibin

Knight

Muttukrishna²

1994

Journal of Endocrinology

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Increased Activity of the Hypothalamic-Pituitary-Adrenal Axis of Rats Exposed to Alcohol in Utero: Role of Altered Pituitary and Hypothalamic Function

Lee

Schmidt²,

Tilders³

et al. 2000

Molecular and Cellular Neuroscience

103

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Heterologous Expression and Characterization of Indian Sahiwal Cattle (Bos indicus) Alpha Inhibin

Bhardwaj

Nayan

Yadav

et al. 2012

Animal Biotechnology

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Inhibin is a non-steroidal glycoprotein hormone of gonadal origin with major action as negative feedback control of the production of FSH by the anterior pituitary gland. The physiological role of inhibin has led to the development of inhibin immunogens for fertility enhancement in farm animals. It is envisaged that a reduction of endogenous inhibin secretion would increase FSH concentrations and thus offers a potential for increasing the number of ovulatory follicles in the ovary. The present work was carried out to produce recombinant bovine (Indian Sahiwal Cattle; Bos indicus) alpha inhibin (bINH-α) in E. coli by optimizing its expression and purification in biologically active form and to study its immunological characterization. A bacterial protein expression vector system based on the phage T(5) promoter was used. The bINH-α encoding gene was successfully cloned and expressed in E. coli and the purified recombinant bINH-α was characterized. Recombinant bINH-α (25 µg mL(-1)) immunized guinea pigs had a significant increase in litter size compared to the control group. These results indicate a role for recombinant bINH-α as a fecundity vaccine to enhance the ovulation rate and litter size in animals.

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Expression of bovine inhibin β subunit in Escherichia coli

Cited by 3 publications

References 33 publications

Measurement of dimeric inhibin using a modified two-site immunoradiometric assay specific for oxidized (Met O) inhibin

Measurement of dimeric inhibin using a modified two-site immunoradiometric assay specific for oxidized (Met O) inhibin

Increased Activity of the Hypothalamic-Pituitary-Adrenal Axis of Rats Exposed to Alcohol in Utero: Role of Altered Pituitary and Hypothalamic Function

Heterologous Expression and Characterization of Indian Sahiwal Cattle (Bos indicus) Alpha Inhibin

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