Skip to main content

Advertisement

SpringerLink
Log in

Determination of alkaline phosphatase isoenzymes and isoforms in dog serum by a simple anion exchange chromatographic method

  • Original Article
  • Published:
Comparative Clinical Pathology Aims and scope Submit manuscript

Abstract

The well-known marker of bone formation is bone alkaline phosphatase, which is produced by osteoblasts and is required for osteoid formation and matrix mineralization. Alkaline phosphatase (ALP, EC 3.1.3.1) is a zinc-containing glycoprotein enzyme catalyzing the hydrolysis of phosphomonoesters at alkaline pHs. ALP is synthesized in a variety of tissues and has different isoenzymes and isoforms. Four variants of ALP have been identified in domestic animals: bone ALP, intestinal ALP, liver ALP, and, in dogs, corticosteroid-induced ALP. Efforts have been made to develop analytical techniques that can differentiate between the different isoenzymes of ALP in animals. These methods include heat inactivation, gel electrophoresis, precipitation with wheat-germ lectin, and, recently, radioimmunoassay and enzyme-linked immunosorbent assay. We developed a simple, fast, and less expensive chromatographic method for differential determination of ALP isoenzymes and isoforms in serum samples. Three peaks of ALP activity were identified in chromatographic fractions. These were related to bone, liver, and steroid isoenzymes of ALP.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Allen MJ (2003) Biochemical markers of bone metabolism in animals: uses and limitations. Vet Clin Pathol 32:101–113

    Article  PubMed  CAS  Google Scholar 

  • Allen MJ, Hoffmann WE, Richardson DC, Breur GJ (1998) Serum markers of bone metabolism in dogs. Am J Vet Res 59:250–254

    PubMed  CAS  Google Scholar 

  • Allen LC, Allen MJ, Breur GJ, Hoffmann WE, Richardson DC (2000a) A comparison of two techniques for the determination of serum bone-specific alkaline phosphatase activity in dogs. Res Vet Sci 68:231–235

    Article  CAS  Google Scholar 

  • Allen MJ, Allen LC, Hoffman WE, Richardson DC, Breur G (2000b) Urinary markers of type I collagen degradation in the dog. Res Vet Sci 69:123–127

    Article  CAS  Google Scholar 

  • Amacher DE, Higgins CV, Schomaker SJ, Clay RJ (1989) Further characterization of serum alkaline phosphatase from male and female beagle dogs. Enzyme 42:1–7

    PubMed  CAS  Google Scholar 

  • Breur GJ, Allen MJ, Carlson SJ, Richardson DC (2004) Markers of bone metabolism in dog breeds of different size. Res Vet Sci 76:53–55

    Article  PubMed  CAS  Google Scholar 

  • Calvo MS, Eyre DR, Gundberg CM (1996) Molecular basis and clinical application of biological markers of bone turnover. Endocr Rev 17:333–368

    PubMed  CAS  Google Scholar 

  • Christgau S, Jensen BO, Bjarnason HN, Henriksen GE, Qvist P, Alexandersen P, Henriksen BD (2000) Serum CrossLaps for monitoring the response in individuals undergoing antiresorptive therapy. Bone 26:505–511

    Article  PubMed  CAS  Google Scholar 

  • Crofton PM (1982) Biochemistry of alkaline phosphatase isoenzymes. Crit Rev Clin Lab Sci 16:161–194

    Article  PubMed  CAS  Google Scholar 

  • DeLaurier A, Jackson B, Ingham K, Pfeiffer D, Horton MA, Price JS (2002) Biochemical markers of bone turnover in the domestic cat: relationships with age and feline osteoclastic resorptive lesions. J Nutr 132:1742S–1744S

    PubMed  CAS  Google Scholar 

  • DeLaurier A, Jackson B, Pfeiffer D, Ingham K, Norton MA, Price JS (2004) A comparison of methods for measuring serum and urinary markers of bone metabolism in cats. Res Vet Sci 77:29–39

    Article  PubMed  CAS  Google Scholar 

  • Dziedziejko V, Safranow K, Zÿyaka DS, Mokrzynska AM, Millo B, Machoya Z, Chlubeka D (2005) Comparison of rat and human alkaline phosphatase isoenzymes and isoforms using HPLC and electrophoresis. Biochim Biophys Acta 1752:26–33

    PubMed  CAS  Google Scholar 

  • Ehrhart N, Dernell WS, Hoffmann WE, Weigel RM, Powers BE, Withrow SJ (1998) Prognostic importance of alkaline phosphatase activity in serum from dogs with appendicular osteosarcoma: 75 cases (1990–1996). J Am Vet Med Assoc 213:1002–1006

    PubMed  CAS  Google Scholar 

  • Fox DB, Cook JL (2001) Synovial fluid markers of osteoarthritis in dogs. J Am Vet Med Assoc 219:756–761

    Article  PubMed  CAS  Google Scholar 

  • Francis DA, Millis DL (2002) Trends observed in bone metabolism markers from dogs following radial ostectomy. Vet Comp Orthop Traumatol 2:A5

    Google Scholar 

  • Gallager JC, Kinyamu HK, Fowler SE, Hughes DB, Dalsky GP, Sherman SS (1998) Calcitrophic hormones and bone markers in the elderly. J Bone Miner Res 13:475–482

    Article  Google Scholar 

  • Garzoto CK, Berg J, Hoffman WE, Rand WM (2000) Prognostic significance of serum alkaline phosphatase activity. Vet Intern Med 14:587–592

    Article  Google Scholar 

  • Goldstein DG, Rogers CE, Harris H (1980) Expression of alkaline phosphatase loci in mammalian tissues. Proc Natl Acad Sci U S A 77:2857–2860

    Article  PubMed  CAS  Google Scholar 

  • Henthorn PS (1996) Alkaline phosphatase. In: Bilezikian JP, Raisz LG, Rodan GA (eds) Principles of bone biology, 1st edn. Academic, San Diego, pp 197–206

    Google Scholar 

  • Hotchkiss CE, Brommage R (2000) Changes in bone turnover during the menstrual cycle in cynomolgus monkeys. Calcif Tissue Int 66:224–228

    Article  PubMed  CAS  Google Scholar 

  • Jackson B, Eastell R, Russel RG, Lanyon LE, Price JS (1996) Measurement of bone-specific alkaline phosphatase in the horse; a comparison of two techniques. Res Vet Sci 61:160–164

    Article  PubMed  CAS  Google Scholar 

  • Kidney BA, Jackson ML (1988) Diagnostic values of alkaline phosphatase isoenzymes separation by affinity chromatography in dogs. Can J Vet Res 52:110–116

    Google Scholar 

  • Komnenou A, Karayannopoulou M, Polizopoulou ZS, Constantinidis TC, Dessiris A (2005) Correlation of serum alkaline phosphatase activity with the healing process of long bone fractures in dogs. Vet Clin Path 34:35–38

    Article  CAS  Google Scholar 

  • Ladlow JF, Hoffmann WE, Breur GJ, Richardson DC, Allen MJ (2002) Biological variability in serum and urinary indices of bone formation and resorption in dogs. Calcif Tissue Int 70:186–193

    Article  PubMed  CAS  Google Scholar 

  • Lammens J, Lui Z, Aerssens J, Dequeker J, Fabry G (1998) Distraction bone healing versus osteotomy healing: a comparative biochemical analysis. J Bone Miner Res 13:279–286

    Article  PubMed  CAS  Google Scholar 

  • Lepage OM, DesCotaux L, Marcoux M, Tremblay A (1991) Circadian rhythms of osteocalcin in equine serum. Correlation with alkaline phosphatase, calcium, phosphate and total protein levels. Can J Vet Res 55:5–10

    PubMed  CAS  Google Scholar 

  • Magnusson P, Lofman O, Larsson L (1992) Determination of alkaline phosphatase isoenzymes in serum by high-performance liquid chromatography with post-column reaction detection. J Chromatogr 576:79–86

    Article  PubMed  CAS  Google Scholar 

  • Nakayama M, Gorai I, Minaguchi H, Rosenquist C, Qvist P (1998) Purification and characterization of bone-specific alkaline phosphatase from a human osteosarcoma cell line. Calcif Tissue Int 62:67–73

    Article  PubMed  CAS  Google Scholar 

  • Paskalev M, Krastev S, Filipov J (2005) Changes in some serum bone markers after experimental fracture and intramedullary osteosynthesis in dogs. Trakia J Sci 3:46–50

    Google Scholar 

  • Philipov JP, Pascalev MD, Aminkov BY, Grosev CD (1995) Changes in serum carboxy terminal telopeptide of type I collagen in an experimental model of canine osteomyelitis. Calcif Tissue Int 57:152–154

    Article  PubMed  CAS  Google Scholar 

  • Price CP (1993) Multiple forms of human serum alkaline phosphatase: detection and quantitation. Ann Clin Biochem 30:355–372

    PubMed  Google Scholar 

  • Price JS, Jackson B, Eastell R, Goodship AE, Blumsohn A, Wright I, Stonham S, Lanyon LE, Russell RGG (1995) Age related changes in biochemical markers of bone metabolism in horses. Equine Vet J 27:201–207

    Article  PubMed  CAS  Google Scholar 

  • Sabellek WM (1988) Alkaline phosphatase laboratory and clinical implications. J Chromatogr 429:419–444

    Article  Google Scholar 

  • Saini PK, Saini SK (1978) Origin of serum alkaline phosphatase in the dog. Am J Vet Res 39:1510–1513

    PubMed  CAS  Google Scholar 

  • Sanecki RK, Hoffmann WE, Gelberg HB, Dorner JL (1987) Subcellular location of corticosteroid-induced alkaline phosphatase in canine hepatocytes. Vet Pathol 24:296–301

    PubMed  CAS  Google Scholar 

  • Sanecki RK, Hoffmann WE, Hansen R, Schaeffer DJ (1993) Quantification of bone alkaline phosphatase in canine serum. Vet Clin Pathol 22:17–23

    Article  PubMed  Google Scholar 

  • Syakalima M, Takiguchi M, Yasuda J, Hashimoto A (1997) The age dependent levels of serum ALP isoenzymes and the diagnostic significance of corticosteroid-induced ALP during long-term glucocorticoid treatment. J Vet Med Sci 59:905–909

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

This research was financially supported by grant no. 86-GR-VT-11 from the Shiraz University Research Council.

Author information

Authors and Affiliations

  1. Department of Biochemistry, School of Veterinary Medicine, Shiraz University, Shiraz, 71345-1731, Iran

    H. R. Shahbazkia & M. Aminlari

  2. Department of Clinical Sciences, School of Veterinary Medicine, Shahrekord University, Shahrekord, Iran

    A. R. Mohamadnia

Authors
  1. H. R. Shahbazkia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Aminlari
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. R. Mohamadnia
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. Aminlari.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shahbazkia, H.R., Aminlari, M. & Mohamadnia, A.R. Determination of alkaline phosphatase isoenzymes and isoforms in dog serum by a simple anion exchange chromatographic method. Comp Clin Pathol 18, 427–432 (2009). https://doi.org/10.1007/s00580-009-0832-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00580-009-0832-9

Keywords

Search

Navigation