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Application of artificial gel antibodies for investigating molecular polymorphisms of human pituitary growth hormone

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Abstract

Artificial gel antibodies were used to investigate human growth hormone (GH) activity in preparations purified from human pituitary glands. A partially purified fraction containing differently sized structural variants of GH was processed to yield monomeric and dimeric forms suitable for synthesizing artificial polyacrylamide gel antibodies. These two types of GH antibodies were used for investigating GH activity in experiments using HPLC gel-permeation and ion-exchange chromatography. In the size-exclusion experiments, both hormone fractions eluted as homogeneous peaks, whereas the ion exchanger resolved the hormones into several active components. The GH monomer antibodies exhibited a much higher affinity for monomeric GH than for dimeric GH, and the GH dimer antibodies exhibited a much higher affinity for dimeric GH than for monomeric GH. It was concluded that these two sets of antibodies might be useful for discriminating between dimeric and monomeric GH in clinical samples.

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Abbreviations

GH:

Growth hormone

TEMED:

N,N,N′,N′-tetramethylethylenediamine

T-unit:

Tiselius-unit (10−5 cm2/V s)

Tris:

Tris(hydroxymethyl)aminomethane

RIA:

Radioimmunoassay

HPLC:

High performance liquid chromatography

CSF:

Human cerebrospinal fluid

ALS:

Amyotrophic lateral sclerosis

References

  • Bagdasarian A, Lahiri B, Talamo RC, Wong P, Colman RW (1974) Brain isoenzyme of creatine kinase. I. Purification of rabbit enzyme and production of specific antibodies. J Clin Invest 54(6):1444–1454

    Article  PubMed  CAS  Google Scholar 

  • Baumann G (1991) Growth hormone heterogeneity: genes, isohormones, variants, and binding proteins. Endocr Rev 12(4):424–449 Review

    Article  PubMed  CAS  Google Scholar 

  • Baumann GP (2009) Growth hormone isoforms. Growth Horm IGF Res 19(4):333–340 (Epub 2009 May 24)

    Article  PubMed  CAS  Google Scholar 

  • Bidlingmaier M (2008) Problems with GH assays and strategies towards standardization. Eur J Endocrinol 159(Suppl 1):S41–S44

    Article  PubMed  CAS  Google Scholar 

  • Brostedt P, Roos P (1988) Isolation of four isomers of the 20,000 dalton variant of human pituitary growth hormone. Prep Biochem 18(3):277–291

    Article  PubMed  CAS  Google Scholar 

  • Brostedt P, Roos P (1989) Isolation of dimeric forms of human pituitary growth hormone. Prep Biochem 19(3):217–229

    Article  PubMed  CAS  Google Scholar 

  • Brostedt P, Luthman M, Wide L, Werner S, Roos P (1990) Characterization of dimeric forms of human pituitary growth hormone by bioassay, radioreceptor assay, and radioimmunoassay. Acta Endocrinol (Copenh) 122(2):241–248

    CAS  Google Scholar 

  • De Palo E, Gatti R, Sicolo Na, De Palo C (1990) Human growth hormone association and binding study in vitro: high performance size exclusion chromatography and radioimmunoassay-measured levels in plasma of normal and acromegalic subjects. Clin Physiol Biochem 8(4):211–220

    PubMed  Google Scholar 

  • De Palo EF, De Filippis V, Gatti R, Spinella P (2006) Growth hormone isoforms and segments/fragments: molecular structure and laboratory measurement. Clin Chim Acta 364(1–2):67–76

    PubMed  Google Scholar 

  • García-Barros M, Costoya JA, Ríos R, Arce V, Devesa J (2000) N-glycosylated variants of growth hormone in human pituitary extracts. Horm Res 53(1):40–45

    Article  PubMed  Google Scholar 

  • Ghasemzadeh N, Nyberg F, Hjertén S (2008a) Highly selective artificial gel antibodies for detection and quantification of biomarkers in clinical samples. I. Spectrophotometric approach to design the calibration curve for the quantification. J Sep Sci 31(22):3945–3953

    Article  PubMed  CAS  Google Scholar 

  • Ghasemzadeh N, Nyberg F, Hjertén S (2008b) Highly selective artificial gel antibodies for detection and quantification of biomarkers in clinical samples. II. Albumin in body fluids of patients with neurological disorders. J Sep Sci 31(22):3954–3958

    Article  PubMed  CAS  Google Scholar 

  • Ghasemzadeh N, Wilhelmsen TW, Nyberg F, Hjertèn S (2010) Precautions to improve the accuracy of quantitative determinations of biomarkers in clinical diagnostics. Electrophoresis 31(16):2722–2729

    Article  PubMed  CAS  Google Scholar 

  • Liao JL, Wang Y, Hjertén S (1996) A novel support with artificially created recognition for the selective removal of proteins and for affinity chromatography. Chromatographia 42:259–262

    Article  CAS  Google Scholar 

  • Ochoa R, Mason M, Fonseca E, Morán C, Zárate A (1993) Distribution of growth hormone isoforms in sera from women with normal ovarian function, galactorrhea, and normoprolactinemia. Fertil Steril 60(2):272–275

    PubMed  CAS  Google Scholar 

  • Oosterom R, Lamberts SW (1985) Comparison of RIA kits with polyclonal antiserum and IRMA kits with monoclonal antibodies for the determination of plasma prolactin and somatotropin in patients with pituitary adenoma. J Clin Chem Clin Biochem 23(1):51–54

    PubMed  CAS  Google Scholar 

  • Roos P, Fevold HR, Gemzell CA (1963) Preparation of human growth hormone by gel filtration. Biochim Biophys Acta 13(74):525–531

    Article  Google Scholar 

  • Roos P, Nyberg F, Brostedt P, Jansson JO, Isaksson O (1987) Isolation of three electrophoretic variants of rat pituitary growth hormone. Prep Biochem 17(1):25–49

    Article  PubMed  CAS  Google Scholar 

  • Silberring J, Brostedt P, Ingvast A, Nyberg F (1991) Analysis of human pituitary growth hormone and its charge variants by fast-atom bombardment mass spectrometry. Rapid Commun Mass Spectrom 5(12):579–581

    Article  PubMed  CAS  Google Scholar 

  • Sinha YN, Lewis UJ (1986) A lectin-binding immunoassay indicates a possible glycosylated growth hormone in the human pituitary gland. Biochem Biophys Res Commun 140:491–497

    Article  PubMed  CAS  Google Scholar 

  • Takátsy A, Sedzik J, Kilár F, Hjertén S (2006) Universal method for synthesis of artificial gel antibodies by the imprinting approach combined with a unique electrophoresis technique for detection of minute structural differences of proteins, viruses, and cells (bacteria): II. Gel antibodies against virus (Semliki Forest Virus). J Sep Sci 29(18):2810–2815

    Article  PubMed  Google Scholar 

  • Wallis M, Ivanyi J, Surowy TK (1982) Binding specificity of monoclonal antibodies towards the size variants of human growth hormone. Mol Cell Endocrinol 28(3):363–372

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This study was supported by grants from the Swedish Research Council (Grant 9459).

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Authors and Affiliations

  1. Department of Pharmaceutical Biosciences, Uppsala University, P.O. Box 591, 751 24, Uppsala, Sweden

    Nasim Ghasemzadeh, Uwe L. Rossbach, Britt-Marie Johansson & Fred Nyberg

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  1. Nasim Ghasemzadeh
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  2. Uwe L. Rossbach
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  3. Britt-Marie Johansson
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  4. Fred Nyberg
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Correspondence to Fred Nyberg.

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Ghasemzadeh, N., Rossbach, U.L., Johansson, BM. et al. Application of artificial gel antibodies for investigating molecular polymorphisms of human pituitary growth hormone. Amino Acids 40, 1249–1255 (2011). https://doi.org/10.1007/s00726-011-0840-3

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  • DOI: https://doi.org/10.1007/s00726-011-0840-3

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