Direct measurement of creatine kinase-MB activity in serum after extraction with a monoclonal antibody specific to the MB isoenzyme.

Abstract Fusion of splenocytes from A/J mice immunized by creatine kinase (EC 2.7.3.2)-MB isoenzyme (CK-MB) with SP2/0-Ag14 myeloma cell line generated hybridomas producing monoclonal antibodies specific to CK-MB. One of these monoclonal antibodies ("Conan-MB") was used to develop a direct assay for CK-MB activity. In the assay, serum is incubated for 30 min at room temperature with "Conan-MB" immobilized on latex beads. The beads are then washed, and CK-MB activity bound to the antibody is measured after incubation with CK enzyme reagent for 30 min at 37 degrees C. Results with the assay correlated well (r = 0.997) with those for CK-MB concentration as measured by a two-site immunoassay. Neither CK-MM, CK-BB, mitochondrial CK, nor a hemolysate of erythrocytes interfered. Use of this unique monoclonal antibody allows rapid, precise, and direct determination of CK-MB activity.

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A New Method for Direct Determination of the Recoilless Fraction with Application to Magnetic Nanoparticles

MRS Proceedings ◽

10.1557/proc-721-e3.7 ◽

2002 ◽

Vol 721 ◽

Author(s):

Monica Sorescu

Keyword(s):

Room Temperature ◽

Average Particle Size ◽

Direct Determination ◽

Average Particle ◽

Lattice Method ◽

Recoilless Fraction ◽

Single Room ◽

Magnetite Particles ◽

Physical Mixtures

AbstractWe propose a two-lattice method for direct determination of the recoilless fraction using a single room-temperature transmission Mössbauer measurement. The method is first demonstrated for the case of iron and metallic glass two-foil system and is next generalized for the case of physical mixtures of two powders. We further apply this method to determine the recoilless fraction of hematite and magnetite particles. Finally, we provide direct measurement of the recoilless fraction in nanohematite and nanomagnetite with an average particle size of 19 nm.

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Preparation of Hybridomas Producing Monoclonal Antibodies against Aflatoxin B1 as a Tool to Control Hepatocellular Carcinoma

International Journal of Pharmacology Phytochemistry and Ethnomedicine ◽

10.18052/www.scipress.com/ijppe.13.1 ◽

2019 ◽

Vol 13 ◽

pp. 1-12

Author(s):

Manal M.E. Ahmed ◽

Rafik Soliman ◽

Jakeen Eljakee ◽

Ahmed El-Sanousi ◽

Haitham Amer ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Aflatoxin B1 ◽

Quantitative Measurement ◽

Myeloma Cell ◽

Selective Medium ◽

Immunization Schedule ◽

Spleen Cells ◽

Myeloma Cell Line ◽

High Affinity ◽

Multiple Uses

Hybridomas that secreted antibodies against aflatoxin B1 for multiple uses were prepared using a unique immunization schedule. Aflatoxin B1-BSA conjugate was used for immunization of Balb/c mice. Spleen cells were harvested from the hyper immunized mice to be fused with myeloma cell line (P3NS1) using polyethylene glycol 3000, 50% concentration as a fusogenic agent. The produced hybridomas were selected using HAT selective medium that was replaced by complete HT medium. From the 10thday after fusion, wells that contain colonies of hybridomas covering 30% or greater of the wells surface were screened for production of monoclonal antibodies against aflatoxin B1 using ELISA. 21 hybridomas were found to be reactive to aflatoxin B1. All were found to belong to IgG2aisotype except one was found to belong to IgM isotype. The prepared monoclonal antibodies and their application to immunoassays represents a useful and rapid quantitative measurement with high affinity and low detection limits in order to purify environmentally occurring levels of this carcinogen specially in areas at high risk for liver cancer.

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Clinical evaluation of an immunoinhibition procedure for creatine kinase-MB.

Clinical Chemistry ◽

10.1093/clinchem/26.1.0150 ◽

1980 ◽

Vol 26 (1) ◽

pp. 150-152

Author(s):

D Obzansky ◽

J A Lott

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Creatine Kinase ◽

Sensitivity And Specificity ◽

Clinical Evaluation ◽

Creatine Kinase Isoenzyme ◽

Creatine Kinase Mb ◽

Diagnostic Sensitivity And Specificity ◽

Creatine Kinase Isoenzyme Mb

Abstract We have clinically evaluated the Dade "Cardiozyme" immunoinhibition procedure for determination of creatine kinase isoenzyme MB (CK-MG) in 71 patients who were suspected of having had an acute myocardial infarction. Electrophoresis for CK-MB was also carried out. On the basis of diagnostic sensitivity and specificity for myocardial infarction, we found the Dade procedure for CK-MB to be somewhat inferior to electrophoresis. In 11 patients for whom the time of infarction was known, we observed normal CK-MB results for two of them by both immunoinhibition and electrophoresis during the first 24 h, but subsequently could detect abnormal CK-MB results by both methods. Thus in some patients such data are not helpful for making a diagnosis in the first 24 h. The Dade procedure is easy to perform, but lacks sensitivity in the region of low CK-MB activity, requires a very stable spectrophotometer, is imprecise, and produces negative numerical results in patients without myocardial infarction.

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Determination of creatine kinase MB activity with the Du Pont aca: interferences from the sample matrix.

Clinical Chemistry ◽

10.1093/clinchem/30.2.238 ◽

1984 ◽

Vol 30 (2) ◽

pp. 238-242 ◽

Cited By ~ 3

Author(s):

W Stein ◽

J Bohner

Keyword(s):

Creatine Kinase ◽

Protein Concentration ◽

Chromatographic Method ◽

Chloride Concentration ◽

Sample Matrix ◽

Creatine Kinase Mb ◽

The Matrix ◽

Enzyme Pattern ◽

Creatine Kinase Isoenzyme Mb

Abstract During the last three years we and other have observed discrepancies between results for creatine kinase isoenzyme MB as measured with the mechanized ion-exchange chromatographic method in the Du Pont aca and those by other techniques. These observations prompted us to investigate the influence of the matrix on the Du Pont CK-MB assay. We conclude that, apart from possible interferences by CK-MM, CK-BB, and both types of macro CK, the aca will give apparent CK-MB activities that are directly related to protein concentration and inversely related to sodium chloride concentration. Practical consequences for the routine and emergency laboratories are: no diluted samples are allowed; application is restricted to samples from patients suspected of acute myocardial infarction which show upper-normal total CK activity; and multiple timed samples are run, in order to recognize the typical change in enzyme pattern with time.

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Direct determination of residual Pluronic F-68 in in-process samples from monoclonal antibody preparations by high performance liquid chromatography

Journal of Chromatography A ◽

10.1016/j.chroma.2010.12.073 ◽

2011 ◽

Vol 1218 (15) ◽

pp. 2106-2113 ◽

Cited By ~ 7

Author(s):

Hyo Helen Chung ◽

Chengfeng Zhou ◽

Hui Koon Khor ◽

Jinshu Qiu

Keyword(s):

Monoclonal Antibody ◽

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

High Performance ◽

Direct Determination ◽

Antibody Preparations

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Information induction for predicting acute myocardial infarction.

Clinical Chemistry ◽

10.1093/clinchem/34.10.2031 ◽

1988 ◽

Vol 34 (10) ◽

pp. 2031-2038 ◽

Cited By ~ 16

Author(s):

R A Rudolph ◽

L H Bernstein ◽

J Babb

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Information Theory ◽

Creatine Kinase ◽

Lactate Dehydrogenase ◽

Noisy Channel ◽

Information Uncertainty ◽

Test Results ◽

Creatine Kinase Mb

Abstract We show how to make an unsupervised discrimination of disease and nondisease states by measuring information and using newer notions of inductive reason. We also present a new theory of group-based reference values that is based on measuring information uncertainty. We use data on the isoenzymes creatine kinase-MB (CK-MB) and lactate dehydrogenase-1 (LD1) and on the percentage of LD1 from 101 patients with acute myocardial infarction (AMI) and from 41 patients with suspected, but unfounded, infarction (non-AMI). Calculating the Shannon entropy, a concept from information theory, of the data base allows determination of a difference in entropy values ("effective information"), which determines decision cutoff values that produce binary-base patterns yielding the fewest classification errors. Redundancy in testing is important because it provides the information to approach a goal of errorless discrimination by coding the test results and meeting the conditions of the "Noisy Channel Theorem" of information theory. This redundancy improves the predictive value of diagnosis by isolating the area of equivocation to evident patterns. Results for CK-MB and LD1 are 99% correct in assigning cases to AMI and non-AMI categories; adding %LD1 increases the proportion of errorless binary patterns from 25% to 90%.

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Identification and characterization of an endothelial, cell-specific antigen with a monoclonal antibody

Blood ◽

10.1182/blood.v66.4.816.816 ◽

1985 ◽

Vol 66 (4) ◽

pp. 816-823 ◽

Cited By ~ 12

Author(s):

WM Parks ◽

RD Gingrich ◽

CE Dahle ◽

JC Hoak

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Vascular Endothelium ◽

Umbilical Vein ◽

Polyacrylamide Gel Electrophoresis ◽

Primary Cultures ◽

Specific Antigen ◽

Myeloma Cell ◽

Membrane Components ◽

And Function

Abstract The purpose of these studies was to use monoclonal antibodies to identify and characterize plasma membrane components unique to the vascular endothelium. Our assumption is that such components may perform some of the specialized functions of the endothelium and, by their identification with antibody probes, we may be able to study further their function and structure. Thus, primary cultures of human umbilical vein endothelium were used to immunize mice whose spleen cells were fused with the mouse myeloma cell NS-1. HEC-1 is a monoclonal antibody derived from such a fusion that appears to react with an antigen located only on endothelial cells. The antigen has been characterized by immunoprecipitation and polyacrylamide gel electrophoresis as a glycoprotein with a mol wt of 180,000 daltons under nonreducing conditions and 90,000 daltons under reducing conditions. Despite a close resemblance to a membrane component shown by others to be a receptor for transferrin, several lines of evidence reported in this paper indicate that this is not the function of the HEC-1 antigen. These data show that monoclonal antibodies can be used to identify and characterize membrane components of the vascular endothelium. Moreover, these probes can be used to inquire about the structure and function of the antigen with which they react.

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