PMID: 11334967May 4, 2001Paper

Monoclonal anti-acid-labile subunit oligopeptide antibodies and their use in a two-site immunoassay for ALS measurement in humans

Journal of Immunological Methods
S StadlerC Strasburger

Abstract

Quantification of the acid-labile subunit (ALS) has to date been restricted to immunoassays utilizing polyclonal antibodies. By immunization with N-terminal and C-terminal specific ALS oligopeptides, we generated monoclonal antibodies (mAbs) that target ALS-specific sequences outside the nonspecific leucine-rich repeats in the ALS molecule. For mAb selection, a special screening method was developed. Monoclonal antibody 5C9, which targets the N-terminus of ALS, is immobilized and the anti-ALS mAb 7H3, directed against the C-terminus, is biotinylated and used as tracer Ab. Due to the extreme pH-lability of ALS, changes in immunorecognition of ALS were investigated after acidification for protein unfolding in different pH ranges and in a time-dependent manner. It was determined that acidification of the serum samples to pH 2.7 for 30 min, followed by neutralization and dilution to 1:100 was the optimal acid-neutralization method. For standardization purposes, a serum pool derived from healthy volunteers was assigned the value 1 U/ml ALS. The sandwich assay has a working range with a linear dose-response curve in a log/log system between 0.005 and 10 U/ml. ALS levels in seven acromegalic patients ranged from 2.0 to 4.2 U/ml, and i...Continue Reading

References

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Citations

Aug 28, 2007·Growth Hormone & IGF Research : Official Journal of the Growth Hormone Research Society and the International IGF Research Society·David L HwangPinchas Cohen
Oct 16, 2007·Growth Hormone & IGF Research : Official Journal of the Growth Hormone Research Society and the International IGF Research Society·Stefan WeissGerald Zimmermann
May 24, 2018·European Journal of Endocrinology·Peter J TrainerMartin Bidlingmaier
Jul 1, 2008·Annals of Indian Academy of Neurology·Angela Vincent
Mar 1, 2007·The Journal of Clinical Endocrinology and Metabolism·Robert D MurrayStephen M Shalet
Dec 31, 2020·The Journal of Clinical Endocrinology and Metabolism·Uta FerrariMichael Drey
Aug 29, 2021·The Journal of Clinical Endocrinology and Metabolism·Emily CottrellHelen L Storr
Sep 22, 2021·BMC Musculoskeletal Disorders·Stefanie JarmuschMichael Drey

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