The influence of the label on the quality of a solid-phase immunoassay: evaluation of a commercial ELISA kit for serum ferritin. 1983

H A Assink, and H J Brouwer, and B G Blijenberg, and B Leijnse

A new Enzyme Linked Immuno Sorbent Assay (ELISA) kit for the determination of serum ferritin has been compared with another ferritin kit based on the Immuno Radio-Metric Assay (IRMA) approach, both assays containing similar antibodies. Based on these studies, we found the within-run precision of the ELISA (and IRMA) to have coefficients of variation of 4-10% and 2-6% respectively, over a concentration range of 12-600 micrograms/l. The between-run precision for the same concentration range exhibited a CV range of 9-13% and 7-11% respectively. The sensitivities were found to be 1.4 micrograms/l and 0.9 microgram/l. The mean recovery was 103% for the ELISA procedure. It was found that, using the serum dilution technique, the linearity reached to 1000 micrograms/l. In the ELISA procedure no influence from the so-called "high dose hook effect" was observed. While EDTA-plasma produced 6% lower values than serum in the ELISA technique, no interference from albumin, gamma-globulins and mild haemolysis was observed. Stability problems with the ELISA kit were not encountered. A comparative analysis of multiple specimens demonstrated nearly identical values with r = 0.994 and y = 0.87 x1.01. The quality and ease of operation of the ELISA approach compared with other techniques are discussed. In conclusion it is possible to replace a radio-label in an immunoassay with an enzyme-label with the same degree of reliability and other parameters of quality control exhibited by radioimmunoassays.

UI MeSH Term Description Entries
D008832 Microchemistry The development and use of techniques and equipment to study or perform chemical reactions, with small quantities of materials, frequently less than a milligram or a milliliter.
D011786 Quality Control A system for verifying and maintaining a desired level of quality in a product or process by careful planning, use of proper equipment, continued inspection, and corrective action as required. (Random House Unabridged Dictionary, 2d ed) Control, Quality,Controls, Quality,Quality Controls
D011863 Radioimmunoassay Classic quantitative assay for detection of antigen-antibody reactions using a radioactively labeled substance (radioligand) either directly or indirectly to measure the binding of the unlabeled substance to a specific antibody or other receptor system. Non-immunogenic substances (e.g., haptens) can be measured if coupled to larger carrier proteins (e.g., bovine gamma-globulin or human serum albumin) capable of inducing antibody formation. Radioimmunoassays
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D005293 Ferritins Iron-containing proteins that are widely distributed in animals, plants, and microorganisms. Their major function is to store IRON in a nontoxic bioavailable form. Each ferritin molecule consists of ferric iron in a hollow protein shell (APOFERRITINS) made of 24 subunits of various sequences depending on the species and tissue types. Basic Isoferritin,Ferritin,Isoferritin,Isoferritin, Basic
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man

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