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 Localization of protective epitopes of the amino terminus of type 5 streptococcal M protein.

1986 ◽  
Vol 163 (5) ◽  
pp. 1191-1202 ◽  
Author(s):  
J B Dale ◽  
E H Beachey
Keyword(s):  
Human Heart ◽  
High Titer ◽  
Heart Tissue ◽  
M Protein ◽  
Amino Terminus ◽  
Particle Phase ◽  
Streptococcal M Protein ◽  
Elisa Inhibition ◽  
Subsequent Chemical ◽  
Human Heart Tissue

We have used a set of overlapping chemically synthesized peptides representing the amino terminus of type 5 streptococcal M protein to localize protective, as opposed to nonprotective and tissue-crossreactive epitopes that might be appropriate for vaccine formulations. Rabbit antisera raised against SM5(1-35) reacted in high titer with pep M5 by ELISA and opsonized type 5 streptococci. None of the antisera crossreacted with human heart tissue or myosin. Antisera against SM5(26-35) reacted with SM5(1-35) and pep M5 but failed to opsonize type 5 streptococci. Particle-phase ELISA indicated that SM5(26-35) antibodies were directed against nonprotective determinants of pep M5 that were not exposed on the surface of viable organisms. Opsonization and ELISA inhibition assays showed that, of the SM5(1-35) antibodies that reacted with M5, all were inhibited by SM5(14-35), whereas none was inhibited by SM5(26-35), suggesting that the protective epitopes of SM5(1-35) resided between residues 14 and 26. This was confirmed by subsequent chemical synthesis of this region; SM5(14-26) totally inhibited SM5(1-35) antibodies that reacted with pep M5 in ELISA, and completely inhibited opsonization of type 5 streptococci by SM5(1-35) antibodies. SM5(14-26) evoked high titers of type-specific, opsonic antibodies against type 5 streptococci, confirming the protective immunogenicity of this 13-residue peptide of type 5 M protein.

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.

scholarly journals IMMUNOLOGIC RELATION OF STREPTOCOCCAL AND TISSUE ANTIGENS

1964 ◽  
Vol 119 (4) ◽  
pp. 651-666 ◽  
Author(s):  
Melvin H. Kaplan ◽  
Kathryn H. Svec
Keyword(s):  
Human Heart ◽  
Streptococcal Infection ◽  
Heart Tissue ◽  
Antigenic Determinants ◽  
Heat Inactivation ◽  
Prolonged Storage ◽  
Acute Glomerulonephritis ◽  
Streptococcal Antigen ◽  
Reactive Antibody ◽  
Human Heart Tissue

Sera from patients with recent streptococcal infection or non-suppurative sequelae exhibit with variable frequency a precipitin reaction in agar gel with a partially purified streptococcal antigen which has been shown previously to be immunologically related to human heart tissue. This precipitin could be absorbed from sera with normal human heart tissue homogenates but not with homogenates of other organs. Demonstration of this cross-reaction by heart absorption was found dependent both upon the serologic properties of individual sera and the nature or state of purification of the streptococcal product employed as test antigen. Evidence was obtained of a close association of heart-related and non-heart-related antigenic determinants in partially purified preparations of the streptococcal antigen by both gel diffusion and immunoelectrophoresis. On immunoelectrophoretic analysis, cross-reactive antigen exhibited a more rapid mobility toward the anode than M protein. It was destroyed by digestion with trypsin, pepsin, and chymotrypsin. Based on specific absorption tests with a Type 5 and Type 19 strain, the antigen was localized to cell walls and to a lesser extent to cell membranes of these strains. Precipitating activity related to cross-reactive antibody was localized to the immunoglobulin zone in immunoelectrophoresis. Reactive sera showed diminution or loss of serological activity following heat inactivation at 56°C or after prolonged storage at 4°C. Sera containing cross-reactive precipitating antibody exhibited an immunofluorescent reaction with sarcolemma of cardiac myofibers, which was inhibited by streptococcal cross-reactive antigen. By this inhibition test, the immunofluorescent reaction related to cross-reactive antibody could be distinguished from that due to other heart-reactive factors. Antibody to streptococcal cross-reactive antigen defined by precipitation-absorption tests was observed in 24 per cent of patients with recent history of uncomplicated streptococcal infection and in the majority of patients with acute rheumatic fever, rheumatic heart disease, or acute glomerulonephritis. It was observed rarely in patients with non-streptococcal related disease. These data provide evidence that induction of cross-reactive autoantibody to heart in certain individuals is associated with streptococcal infection.

scholarly journals Factors for C-Kit Expression in Cardiac Outgrowth Cells and Human Heart Tissue

2017 ◽  
Vol 58 (6) ◽  
pp. 962-968
Author(s):  
Satoshi Matsushita ◽  
Kazuo Minematsu ◽  
Taira Yamamoto ◽  
Hirotaka Inaba ◽  
Kenji Kuwaki ◽  
...  
Keyword(s):  
Human Heart ◽  
Heart Tissue ◽  
Human Heart Tissue

scholarly journals Chronotropic Responses of Human Heart Tissue Cultures

1972 ◽  
Vol 30 (6) ◽  
pp. 628-633 ◽  
Author(s):  
T. D. CHANG ◽  
G. R. CUMMING
Keyword(s):  
Human Heart ◽  
Heart Tissue ◽  
Tissue Cultures ◽  
Human Heart Tissue
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