Radioimmunoassay of creatine kinase isoenzymes in human serum: isoenzyme BB.

1978 ◽  
Vol 24 (3) ◽  
pp. 422-428 ◽  
Author(s):  
M H Zweig ◽  
A C Van Steirteghem ◽  
A N Schechter
Keyword(s):  
Creatine Kinase ◽  
Human Serum ◽  
Cross Reactivity ◽  
Healthy Adults ◽  
Serum Samples ◽  
Creatine Kinase Isoenzymes ◽  
Specific Radioimmunoassay ◽  
Assay Precision ◽  
Free Antigen ◽  
Healthy Persons

Abstract We describe a sensitive, specific radioimmunoassay for the BB isoenzyme of creatine kinase (CK-BB) in serum. A sequential saturation assay was used to achieve sufficient sensitivity to detect the isoenzyme in 100-microliter serum samples of all healthy persons and patients tested. Bound and free antigen were separated by a second antibody system. Large excesses of purified isoenzyme MM did not react in the assay. Cross reactivity of two preparations of CK-MB was only 1 to 7+. The 95th percentile of serum CK-BB in 208 healthy adults was 6.2 microgram/liter. Within-assay and between-assay precision ranged from 5.5 to 11.9% and 9.7 to 13.6%, respectively.

Radioimmunoassay of creatine kinase isoenzymes in human serum: isoenzyme MM.

1978 ◽  
Vol 24 (3) ◽  
pp. 414-419 ◽  
Author(s):  
A C Van Steirteghem ◽  
M H Zweig ◽  
A N Schechter
Keyword(s):  
Creatine Kinase ◽  
Enzymatic Activity ◽  
Human Serum ◽  
Mass Concentration ◽  
High Titer ◽  
Cross Reactivity ◽  
Serum Enzymes ◽  
Serum Concentrations ◽  
Specific Radioimmunoassay ◽  
Chloramine T

Abstract Measurement of the mass concentration of serum enzymes by radioimmunoassay provides direct quantitation of specific isoenzymes and may be less subject to some of the limitations of traditional assay procedures for enzymes. We describe the development of a sensitive and specific radioimmunoassay for the muscle isoenzyme of creatine kinase, CM-MM, in human serum. CK-MM, purified from human skeletal muscle, was used to raise high-titer antisera and for iodination by the Chloramine T method. The radioimmunoassay required 50 microliter of sample, utilized a double-antibody separation method, and was completed in 24 h. Cross reactivity with CK-BB was virtually zero, 3--17% with CK-MB. The mass concentration of CK-MM in the serum of healthy subjects ranged from 36 to 1668 microgram/liter and correlated closely with total CK enzymatic activity. Serum concentrations of CK-MM from casually selected patients correlated less well with total CK enzymatic activity, suggesting the existence of other CK isoenzymes or the presence of inactive forms.

Creatine kinase isoenzymes of mitochondrial origin in human serum.

1979 ◽  
Vol 25 (6) ◽  
pp. 943-947 ◽  
Author(s):  
G P James ◽  
R L Harrison
Keyword(s):  
Skeletal Muscle ◽  
Creatine Kinase ◽  
Lactate Dehydrogenase ◽  
Serum Samples ◽  
Muscle Creatine Kinase ◽  
Cellulose Acetate Electrophoresis ◽  
University Of Texas ◽  
Creatine Kinase Isoenzymes ◽  
Mitochondrial Origin ◽  
Muscle Creatine

Abstract We measured creatine kinase (EC 2.7.3.2) activity in 1009 serum samples from 538 patients in the intensive-care units of the University of Texas Medical Branch hospitals. Creatine kinase isoenzymes migrating cathodal to skeletal muscle creatine kinase (CK-MM) on cellulose acetate electrophoresis were found in sera from 14 of the 538 patients. Creatine kinase, lactate dehydrogenase (EC 1.1.1.27), aspartate aminotransferase (EC 2.6.1.1), and alanine aminotransferase (EC 2.6.1.2) activities were abnormally increased in these 14 patients. Liver lactate dehydrogenase isoenzyme (LDH5) and cardiac creatine kinase isoenzyme (CK-MB) were abnormally increased in 12 and eight of these patients, respectively. Ten of the 14 patients died during their hospital admission. We believe the creatine kinase isoenzymes that migrated cathodal to skeletal muscle creatine kinase (CK-MM) were of mitochondrial origin.

RADIOIMMUNOASSAY OF 3,3′-DI-IODOTHYRONINE IN UNEXTRACTED SERUM: THE EFFECT OF ENDOGENOUS TRI-IODOTHYRONINE

1978 ◽  
Vol 79 (3) ◽  
pp. 357-362 ◽  
Author(s):  
T. J. VISSER ◽  
L. M. KRIEGER-QUIST ◽  
R. DOCTER ◽  
G. HENNEMANN
Keyword(s):  
Human Serum ◽  
Limit Of Detection ◽  
Cross Reactivity ◽  
Transport Proteins ◽  
Normal Serum ◽  
Serum Concentrations ◽  
Specific Radioimmunoassay ◽  
Highly Sensitive ◽  
The Mean ◽  
Mean Concentration

The development of a highly sensitive and specific radioimmunoassay for 3,3′-di-iodothyronine (3,3′-T2) is described. The assay was applied to the measurement of 3,3′-T2 in unextracted human serum and used 8-anilino-l-naphthalene-sulphonic acid to inhibit the binding of 3,3′-T2 to serum transport proteins. The lower limit of detection of the assay was 2 fmol 3,3′-T2 per tube, which corresponded to 10 pmol 3,3′-T2/l serum. The mean concentration of 3,3′-T2 in normal serum was found to be 23 pmol/l, which is considerably lower than most values reported previously. Evidence is presented which suggests that the cross-reactivity of tri-iodothyronine with the antiserum to 3,3′-T2 is an important factor in the measurement of serum concentrations of 3,3′-T2 by radioimmunoassay.

Variability among commercially available digoxin radioimmunoassay kits in cross reactivity to dihydrodigoxin.

1978 ◽  
Vol 24 (1) ◽  
pp. 155-157 ◽  
Author(s):  
W G Kramer ◽  
N L Kinnear ◽  
H K Morgan
Keyword(s):  
Human Serum ◽  
Therapeutic Range ◽  
Cross Reactivity ◽  
Serum Samples ◽  
Serum Digoxin ◽  
The Cross ◽  
High Degree ◽  
Free Serum ◽  
Digoxin Radioimmunoassay

Abstract We evaluated four commercially available 125I-digoxin radioimmunoassay kits with regard to their ability to cross react with the digoxin metabolite dehydrodigoxin. We prepared dihydrodigoxin serum samples in digoxin-free serum over the concentration range 0.4 to 5.0 microgram/liter and assayed them with each kit according to the manufacturer's instructions. The metabolite was able to displace the 125I-labeled digoxin derivative from the antibody supplied with all four kits. However, the extent of the cross reactivity depended on the kit, ranging from essentially zero to a high degree of interference. Dihydrodigoxin is the only metabolite of digoxin to have been quantitiated in human serum, and may comprise up to 30% of total glycosides. Over the clinical and therapeutic range of serum digoxin concentrations, enough dihydrodigoxin can be produced to interfere in the determination of serum digoxin concentrations by this method. We suggest that laboratories evaluate their specific kit with regard to cross reactivity to this metabolite.

scholarly journals Specificity and Diversity of Antibodies to Mycobacterium tuberculosis Arabinomannan

2003 ◽  
Vol 10 (1) ◽  
pp. 88-94 ◽  
Author(s):  
Josephine Anne D. Navoa ◽  
Suman Laal ◽  
Liise-Anne Pirofski ◽  
Gary R. McLean ◽  
Zhongdong Dai ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Human Serum ◽  
Cross Reactivity ◽  
Human Infection ◽  
Serum Samples ◽  
Purified Protein Derivative ◽  
Human Antibodies ◽  
Human Serum Samples ◽  
Negative Controls ◽  
Murine Mabs

ABSTRACT Arabinomannan (AM) is a polysaccharide antigen of the mycobacterial capsule. However, it is uncertain whether AM constitutes an immunologically distinct fraction of Mycobacterium tuberculosis. In this study, we analyzed the repertoire and specificity of antibodies to AM by using AM-binding murine monoclonal antibodies (MAbs) and human serum samples. Murine MAbs were found to be diverse in their specificity to AM and cross-reactivity with other arabinose-containing mycobacterial polysaccharides, with MAb 9d8 binding exclusively to AM. Human antibodies to AM were detected in serum samples from patients with pulmonary tuberculosis (TB), as well as in those from healthy, purified protein derivative-negative controls, with significantly higher titers among patients. The binding of human antibodies to AM was inhibited by MAb 9d8 in three patients with TB but not in controls. MAb 5c11, which recognizes other mycobacterial arabinose-containing carbohydrates in addition to AM, inhibited the binding of serum samples from 75% of patients and 76% of controls. Analysis of human antibodies with murine MAbs to human VH determinants demonstrated diversity among antibodies to AM with qualitative and quantitative differences compared with antibodies to lipoarabinomannan. In summary, our study suggests that antibodies to AM are diverse and heterogeneous with respect to antigen recognition and VH determinant expression, with human serum samples containing different subsets of antibodies to AM with the specificities of AM-binding murine MAbs. One MAb and a subset of human antibodies bind AM specifically, suggesting that this polysaccharide is antigenically distinct and is expressed in human infection.

Radioimmunoassay of aspartate aminotransferase isoenzymes in human serum.

1984 ◽  
Vol 30 (8) ◽  
pp. 1361-1365 ◽  
Author(s):  
F Y Leung ◽  
A E Niblock ◽  
A R Henderson
Keyword(s):  
Human Serum ◽  
Column Chromatography ◽  
Aspartate Aminotransferase ◽  
Human Heart ◽  
Solid Phase ◽  
High Titer ◽  
Heart Tissue ◽  
Cross Reactivity ◽  
Specific Radioimmunoassay ◽  
Human Heart Tissue

Abstract We describe the development of a sensitive, specific radioimmunoassay for the cytoplasmic and mitochondrial isoenzymes of human aspartate aminotransferase (L-aspartate:2-oxoglutarate aminotransferase; EC 2.6.1.1). Isoenzymes from human heart tissue were purified to homogeneity and used to raise high-titer antisera in rabbits. We partly purified the antisera by selective column chromatography. The Bolton-Hunter reagent was used to radioiodinate the isoenzymes. The assay requires 100 microL of serum, includes a solid-phase second-antibody separation, and can be completed in less than 3 h. There was no cross reactivity between the two isoenzymes. As little as 5 micrograms (50 pmol) of each aspartate aminotransferase can be measured per liter of serum.

scholarly journals Flaxseed Ingestion Alters Ratio of Enterolactone Enantiomers in Human Serum

2010 ◽  
Vol 2010 ◽  
pp. 1-5 ◽  
Author(s):  
Niina M. Saarinen ◽  
Annika I. Smeds ◽  
José L. Peñalvo ◽  
Tarja Nurmi ◽  
Herman Adlercreutz ◽  
...  
Keyword(s):  
Pilot Study ◽  
Human Serum ◽  
Human Subjects ◽  
Healthy Adults ◽  
Major Determinant ◽  
Parent Plant ◽  
Serum Samples ◽  
Time Resolved ◽  
Chromatographic Methods ◽  
Habitual Diet

Enterolactone (EL) is an enterolignan found in human subjects. In this pilot study, the enantiomeric ratios of serum EL were determined in serum from healthy adults during consumption of habitual diet, and after an 8-day supplementation with flaxseed (25 g/day). (−)EL dominated in all serum samples collected during habitual diet consumption. However, the ratio of (−)EL and (+)EL enantiomers differed markedly between individuals. Flaxseed ingestion increased significantly the proportion of (+)EL in all subjects. Moreover, a small but significant increase in serum (−)EL concentration was measured. After flaxseed ingestion, (−)EL concentrations correlated with those of (+)EL suggesting that the stereochemistry of the parent plant lignan in flaxseed is not a major determinant of EL formation in human subjects. Comparison of EL concentrations obtained with the validated chromatographic methods (HPLC-MS/MS, HPLC-CEAD, and GC-MS) and the time-resolved fluoroimmunoassay (TR-FIA) revealed that the immunoassay method underestimates human serum EL concentrations after the flaxseed ingestion.

scholarly journals Lower seroreactivity to European than to North American H3N2 swine influenza viruses in humans, Luxembourg, 2010

2015 ◽  
Vol 20 (13) ◽  
Author(s):  
Y Qiu ◽  
C P Muller ◽  
K Van Reeth
Keyword(s):  
Human Serum ◽  
North American ◽  
Strong Correlation ◽  
Haemagglutination Inhibition ◽  
Swine Influenza ◽  
Cross Reactivity ◽  
Influenza Viruses ◽  
Serum Samples ◽  
Human Serum Samples ◽  
The North

Seroreactivity to H3N2 swine influenza viruses (SIVs) was evaluated in serum samples collected from 843 people aged 0 to 100 years in 2010 in Luxembourg. Sera were analysed by haemagglutination inhibition (HI) and virus neutralisation (VN) assays targeting a European H3N2 SIV, a North American H3N2 variant of swine origin (H3N2v) and human seasonal H3N2 viruses isolated in 1975, 1995 and 2005. HI antibodies (titre?≥?10) against European H3N2 SIV were almost exclusively detected in those born before 1990, of whom 70% were seropositive. HI antibodies against H3N2v were predominantly found in those born before 2000, with 86% seropositive. Titres against the North American H3N2v were higher than against the European H3N2 SIV. VN patterns were similar, but with higher rates and titres. We also demonstrated lower seroreactivity to European H3N2 SIV than to North American H3N2v virus. Finally, we found a strong correlation between HI titres against the European H3N2 SIV and H3N2v and their respective human ancestors, A/Victoria/3/75 and A/Nanchang/933/95. This finding and the minimal contacts between humans and pigs in Luxembourg suggest that anti-SIV antibodies in human serum samples reflect serological cross-reactivity with historical human H3N2 viruses. Our findings help assess the pandemic risk of H3N2 SIV.

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