An interpretation of the serum alkaline phosphatase isoenzyme patterns in patients with obstructive liver disease.

Price, Christopher P.; Sammons, H. G.

doi:10.1136/jcp.29.11.976

Cited by 13 publications

(6 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The selective binding of soluble serum alkaline phosphatase to a monoclonal antibody immunoadsorbent may allow the development of an assay to measure the unbound particulate form of the enzyme in serum. At present there is no simple quantitative method of doing this, yet it has been proposed that the presence of the high-Mr particulate form of alkaline phosphatase in serum is a good indicator of obstructive liver disease (Price & Sammons, 1976;Siede & Seiffert, 1983).…”

Section: Discussionmentioning

confidence: 99%

“…Part of the serum alkaline phosphatase of patients with hepatobiliary disease is of high Mr and may be complexed with other plasma membrane enzymes and lipids. High-Mr alkaline phosphatase is diagnostic of obstructive liver diseases, particularly extra-hepatic obstruction (De Broe et al, 1975;Price & Sammons, 1976;Moss, 1982;Siede & Seiffert, 1983). The origin of high-M, alkaline phosphatase and its relationship with soluble serum enzyme has generated considerable controversy (Price & Sammons, 1974;Crofton & Smith, 1981;De Broe et al, 1985;Kaplan, 1986;Wulkan & Leijnse, 1986;Maguire et al, 1986).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The preparation of monoclonal antibodies to human bone and liver alkaline phosphatase and their use in immunoaffinity purification and in studying these enzymes when present in serum

Bailyes

Seabrook

Calvin

et al. 1987

Biochemical Journal

Self Cite

View full text Add to dashboard Cite

1. Liver and bone alkaline phosphatase isoenzymes were solubilized with the zwitterionic detergent sulphobetaine 14, and purified to homogeneity by using a monoclonal antibody previously raised against a partially-purified preparation of the liver isoenzyme. Both purified isoenzymes had a specific activity in the range 1100-1400 mumol/min per mg of protein with a subunit Mr of 80,000 determined by SDS/polyacrylamide gel electrophoresis. Butanol extraction instead of detergent solubilization, before immunoaffinity purification of the liver enzyme, resulted in the same specific activity and subunit Mr. The native Mr of the sulphobetaine 14-solubilized enzyme was consistent with the enzyme being a dimer of two identical subunits and was higher than that of the butanol-extracted enzyme, presumably due to the binding of the detergent micelle. 2. Pure bone and liver alkaline phosphatase were used to raise further antibodies to the two isoenzymes. Altogether, 27 antibody-producing cell lines were cloned from 12 mice. Several of these antibodies showed a greater than 2-fold preference for bone alkaline phosphatase in the binding assay used for screening. No antibodies showing a preference for liver alkaline phosphatase were successfully cloned. None of the antibodies showed significant cross-reaction with placental or intestinal alkaline phosphatase. Epitope analysis of the 27 antibodies using liver alkaline phosphatase as antigen gave rise to six groupings, with four antibodies unclassified. The six major epitope groups were also observed using bone alkaline phosphatase as antigen. 3. Serum from patients with cholestasis contains soluble and particulate forms of alkaline phosphatase. The soluble serum enzyme had the same size and charge as butanol-extracted liver enzyme on native polyacrylamide-gel electrophoresis. Cellulose acetate electrophoresis separated the soluble and particulate serum alkaline phosphatases as slow- and fast-moving forms respectively. In the presence of sulphobetaine 14 all the serum enzyme migrated as the slow-moving form on cellulose acetate electrophoresis. Monoclonal anti-(alkaline phosphatase) immunoadsorbents did not bind the particulate form of alkaline phosphatase in cholestatic serum but bound the soluble form. In the presence of sulphobetaine 14 all the cholestatic serum alkaline phosphatase bound to the immunoadsorbents. 4. The electrophoretic and immunological data are consistent with both particulate and soluble forms of alkaline phosphatase in cholestatic serum being derived from the hepatocyte membrane.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The preparation of monoclonal antibodies to human bone and liver alkaline phosphatase and their use in immunoaffinity purification and in studying these enzymes when present in serum

Bailyes

Seabrook

Calvin

et al. 1987

Biochemical Journal

Self Cite

View full text Add to dashboard Cite

show abstract

“…Alkaline phosphatase, one of the enzymes that hydrolyze phosphate compounds in alkaline pH environments, is present in all tissues throughout the body, especially in the liver, bile duct, bone, placenta, and intestinal mucosa. Increased serum alkaline phosphatase levels have been found in patients with cancer with liver and bone metastasis ( 18 ), as well as in those with liver diseases, biliary obstruction, and pregnancy and among children ( 23 , 24 ). Regarding the involvement of alkaline phosphatase in cryptogenic stroke, the effects of alkaline phosphatase on blood vessel calcification and arteriosclerosis have been reported ( 25 ), as has the association between high serum alkaline phosphatase levels and poor functional outcomes in patients with ischemic stroke, due to the previously considered mechanisms of inflammation, malnutrition, and infection that can occur in active cancer, which has been reported to be associated with elevated alkaline phosphatase levels ( 26 , 27 ).…”

Section: Discussionmentioning

confidence: 99%

Serum Alkaline Phosphatase in Cryptogenic Stroke Cases with Active Cancer

et al. 2022

View full text Add to dashboard Cite

Objective We assessed the relationship between the levels of serum alkaline phosphatase, which is often increased with biliary obstruction and bone metastasis, and active cancer in patients with cryptogenic stroke. Methods Serum alkaline phosphatase levels in patients with cryptogenic stroke sampled upon admission were measured using the Japan Society of Clinical Chemistry method used in Japan. Active cancer was defined as a new diagnosis, treatment, progression, or recurrence within six months before admission or metastatic cancer. Multivariate logistic regression analyses were performed to explore the relationship between serum alkaline phosphatase and active cancer in these patients. Results Among the 249 patients classified as having cryptogenic stroke, 64 had active cancer. Patients with cryptogenic stroke with active cancer had significantly higher serum alkaline phosphatase levels (486±497 vs. 259±88.2 U/L; p<0.001) than those without cancer. Multivariate logistic analysis revealed that serum alkaline phosphatase levels ≥286 U/L were associated with cryptogenic stroke with active cancer [odds ratio (OR), 2.669, 95% confidence interval (CI), 1.291-5.517; p=0.008] independent of age ≤70 years old (OR, 3.303, 95% CI, 1.569-6.994; p=0.002), male sex (OR, 0.806, 95% CI, 0.380-1.710; p=0.573), and serum D-dimer levels ≥2.6 μg/mL (OR, 18.78, 95% CI, 8.130-43.40; p<0.001). Conclusion In patients with cryptogenic stroke, high serum alkaline phosphatase levels may be related to active cancer.

show abstract

“…The origin (biliary) isoenzyme for both ALP and yGT has been shown to be increased in obstructive liver disease (Price and Sammons, 1976;Wenham et al, 1979). The studies presented above therefore allow a quantitative assay to be made of this isoenzyme.…”

Section: Meanmentioning

confidence: 98%

Quantitative Recovery of Alkaline Phosphatase and γ-Glutamyl Transferase Isoenzymes after Polyacrylamide Gel Electrophoresis

Brooks¹,

Sammons²

1980

Ann Clin Biochem

View full text Add to dashboard Cite

SUMMARY Loss of activity of the isoenzymes of v-glutamyl transferase and alkaline phosphatase has been shown to occur during electrophoresis on polyacrylamide gel. After studying the possible factors concerned in this loss, reasonable recovery from the gel can be obtained only for the isoenzyme staying at the origin. Maximum recovery is 60% for origin y-glutamyl transferase and 92 % for origin alkaline phosphatase.Recent work from this department indicates that although the isoenzymes of y-glutamyl transferase (yGT) are well separated when using polyacrylamide gel electrophoresis, and easily identified by staining, the actual recovery is only 30 % of the original activity. As this is a most effective and clinically useful medium for separating not only the isoenzymes of yGT (Azzopardi and Jayle, 1973) but also those of alkaline phosphatase (ALP) (Walker and Pollard, 1971;Price and Sammons, 1974), it seemed desirable to investigate this phenomenon further, firstly, to see if ALP behaved similarly, secondly, to see if the extent of recovery was similar for all the isoenzymes separated, and, thirdly, to identify if possible the factors involved. Methods ELECTROPHORESISShandon disc electrophoresis equipment was used for both enzymes. The tubes were 7·5 ern in length with a bore of 0·5 cm. Gels were prepared for yGT by the method of Azzopardi and Jayle (1973) and for ALP by the method of Smith et al. (1968). These use respectively 7 % and 5 % polyacrylamide made up in Tris-glycine buffer pH 8'3 and Tris-borate buffer pH 9·5for optimum separation of the isoenzyrnes, A 20~l serum sample was applied to the gel after dilution 1:1 with 40 %sucrose. The mobility of the separation was marked by pre-staining the diluted serum with a grain of bromophenol blue. Separation was carried out at 1·0 rnA/gel constant current for 5 minutes to allow for penetration into the gel and then raised to "Present address: Group Pathology Laboratory, Lakin Road, Warwick. 154 rnA/gel until the albumin-bromophenol blue complex reached the bottom of the gel. After electrophoresis the gels were rimmed from the tubes and the enzymes were measured in the appropriate length of gel. Each 0·5 ern had a volume of 0'1 ml and contained 35 mg (for yGT) and 25 mg (for ALP) of polyacrylamide. ENZYME METHODS v-Glutamyl transferaseA manual adaptation of the kinetic yGT assay of Rosalkiand Tarlow (1974) using y-L-glutamylp-nitraniJide as substrate at a pH of 8'0 and at 37°C was used. The reaction was stopped by adding 25 %acetic acid. The change in optical density at 410 nm was then used as a measure of activity. The coefficient of variation of the method was found to be 7·2 %.Alkaline phosphatase A manual modification of the optimised conditions recommended by the Committee on Enzymes (1977) using p-nitrophenyl phosphate was used. After incubation for 30 minutes, the reaction was stopped by the addition of 0·4M NaOH. The optical density was read at 410 nm. Controls were assayed by adding 0'4M NaOH before the addition of substrate. The coefficient of variatio...

show abstract

An interpretation of the serum alkaline phosphatase isoenzyme patterns in patients with obstructive liver disease.

Cited by 13 publications

References 17 publications

The preparation of monoclonal antibodies to human bone and liver alkaline phosphatase and their use in immunoaffinity purification and in studying these enzymes when present in serum

The preparation of monoclonal antibodies to human bone and liver alkaline phosphatase and their use in immunoaffinity purification and in studying these enzymes when present in serum

Serum Alkaline Phosphatase in Cryptogenic Stroke Cases with Active Cancer

Quantitative Recovery of Alkaline Phosphatase and γ-Glutamyl Transferase Isoenzymes after Polyacrylamide Gel Electrophoresis

Contact Info

Product

Resources

About