Enzyme-linked immunosorbent protein

Morishita, N., Kamiya, K., Matsumoto, T., Sakai, S., Teshima, R., Urisu, A., Moriyama, T., Ogawa, T., Akiyama, H., and Morimatsu, F. (2008). A reliable enzyme-linked immunosorbent assay for determination of soybean proteins in processed foods. /. Agric. Food Chem. 56, 6818-6824. 

Additionally it has been our experience that mass spectrometry as a routine detection/identification technique for bacteria is not well received by microbiologists and clinicians who prefer less expensive, less complicated approaches to bacterial typing and identification, such as methods based on polymerase chain reaction (PCR) and enzyme-linked immunosorbent assays (ELISA). For that reason we have adapted our MS approach to serve as a means of biomarker discovery that feeds candidate proteins or leads into development as PCR targets or other immunoassay techniques. 

In order to measure the exact amount of a specific protein (analyte) by IHC signal intensity, a critical requirement is the availability of a standard reference material (present in a known amount by weight) that can be used to calibrate the assay (IHC stain). It is then possible to determine the amount of test analyte (protein) by a translation process from the intensity of IHC signals. In this respect it is helpful to consider the IHC stain as a tissue based ELISA assay (Enzyme Linked ImmunoSorbent Assay), noting that ELISA is used in the clinical laboratory as a standard quantitative method for measuring protein by weight in fluids, by reference to a calibrating reference standard. 

The use of independent methods, other than IFIC, for quantitative demonstration of proteins is particularly important. Both enzyme-linked immunosorbent assay (ELISA) and Western blot may be employed to confirm the amount of protein in a cell/tissue model, and in the protein-embedding bar code model under both comparable fresh and FFPE samples for accurate... 

In the last decade, modem biochemical methods have been used for analysis of protein binders [20,21] in one case a group led by A. Heginbotham identified egg proteins in a seventeenth century painting using immunofluorescent microscopy and enzyme-linked immunosorbent assay (ELISA) [20]. 

A. Heginbotham, V. Millay, M. Quick, The use of immunofluorescence microscopy (IFM) and enzyme linked immunosorbent assay (ELISA) as complementary techniques for protein identi fication in artists materials, J. Am. Inst. Cons., 45, 89 105 (2006). 

Winterbourn, C.C., and Buss, I.H. (1999) Protein carbonyl measurement by enzyme-linked immunosorbent assay. Meth. Enzymol. 300, 106-111. 

A classical approach is the enzyme linked immunosorbent assay (ELISA), where the antigen (e.g., the protein to be quantified) is immobilized on the surface of a well. A first antigen-specific antibody is applied to occupy all antigens, before a second antibody binds all primary antibodies on the well. The second antibody carries an enzyme, which now catalyzes a color reaction. If the substrate of the enzyme is given in high excess, the enzyme is saturated and the production of product is linear with time and concentration of second antibody and antigen (Fig. 8). 

In summary, these are the clinically relevant questions about the immunogenicity of rDNA species-specific proteins will antibody be induced in the recipient that will neutralize the therapeutic effect or lead to immune complex disease What is the class (e.g., IgG or IgE) and specificity (i.e., reactivity against specific protein or contaminant) of the antibody induced The former antibody type could potentially neutralize the product and produce immune complex disease, while the latter could result in an anaphylaxis response. It is possible that the antibody induced is of insignificant health consequence, and its presence is known only because of improvements made in the sensitivity of detection methods with the introduction of the enzyme-linked immunosorbent (ELISA) assay. 

PAMAM dendrimers or modified PAMAM dendrimers (with oxiamine or sulfhydryl surface functionalities) exclusive of one another. These antisera recognize den-drimers in ELISA (enzyme-linked immunosorbent assays), dot blots and Western blots. The immunogenicity of dendrimer-protein conjugates has implications for therapeutic use of dendrimers as vaccines and we anticipate that antidendrimer antibodies will have applications in patterning and assembling nanostructures containing dendrimers. 

Immune detection is a key utility of antibodies in biotechnology [3, 5]. Antiden-drimer sera efficiently detect dendrimers in multiple assay formats, including enzyme-linked immunosorbent assays (ELISA), and in Western and dot blots [3, 5], ELISA assays are commonly used to quantitate proteins, and a quantitative ELISA could be developed for dendrimers using our sera, though doing so would require development of dendrimer standards of known concentration that could be used for calibration. 

D. J. Bucher, A. Mikhail, S. Popple, P. Graves, G. Meiklejohn, D. S. Hodes, K. Johansson, andP. E. Halonen, Rapid detection of Type A Influenza viruses with monoclonal antibodies to the M protein (Ml) by enzyme-linked immunosorbent assay and time-resolved fluoroimmunoassay, J. Clin. Microbiol. 29, 2484-2488 (1991). 

It needs to be mentioned here that there remains some controversy over the placental expression of P-gp as a function of gestational age. An immunohis-tochemical study done by Macfarland et al. showed that P-gp was localized to the microvillous border of trophoblasts in first trimester placenta, but not in term placenta [85], Subsequent studies refuted this to show that MDR1 mRNA is present throughout pregnancy [94], More recently, enzyme-linked immunosorbent assay (ELISA) performed in syncytial microvillous membrane showed that P-gp protein expression in early gestational age placenta is about... 

Braun, A. and J. Alsenz (1997). Development and use of enzyme-linked immunosorbent assays (ELISA) for the detection of protein aggregates in interferon-alpha (IFN-alpha) formulations. Pharm Res 14(10) 1394-1400. 

Lucas, C., C. Nelson, M.L. Peterson, S. Frie, D. Vetterlein, T. Gregory, and A.B. Chen (1988). Enzyme-linked immunosorbent assays (ELISAs) for the determination of contaminants resulting from the immunoaffinity purification of recombinant proteins. J Immunol Methods 113(1) 113-122. 

Engvall E, Jonsson K, Perlmann P. 1971. Enzyme-linked immunosorbent assay. II. Quantitative assay of protein antigen, immunoglobulin G, by means of enzyme-labelled antigen and antibody-coated tubes. Biochim Biophys Acta 251 427-434. 

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