Monoclonal Antibody Detection of Porcine Meat

1994 ◽  
Vol 57 (2) ◽  
pp. 146-149 ◽  
Author(s):  
P. MORALES ◽  
T. GARCÍA ◽  
I. GONZÁLEZ ◽  
R. MARTÍN ◽  
B. SANZ ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Bovine Serum Albumin ◽  
Horseradish Peroxidase ◽  
Indirect Elisa ◽  
Cross Reactivity ◽  
Antibody Detection ◽  
Muscle Proteins ◽  
Elisa Format ◽  
Soya Proteins

A stable hybridoma cell line (DD9) has been produced secreting a monoclonal antibody specific for porcine muscle proteins. The DD9 monoclonal antibody (mAb) failed to show a significant cross-reactivity when tested against beef, horse, chicken, and soya proteins, as well as bovine caseins, gelatin, and bovine serum albumin. The DD9 mAb was further used in an indirect ELISA format for detection of defined amounts of porcine meat (1–100%) in beef and chicken meat mixtures immobilized on 96-well plates. Immunorecognition of monoclonal antibodies adsorbed to porcine meat was made with rabbit anti-mouse immunoglobulins conjugated to the enzyme horseradish peroxidase.

scholarly journals Heterologous screening of hybridomas for the development of broad-specific monoclonal antibodies against deoxynivalenol and its analogues

2014 ◽  
Vol 7 (3) ◽  
pp. 257-265 ◽  
Author(s):  
Y. Guo ◽  
M. Sanders ◽  
A. Galvita ◽  
A. Heyerick ◽  
D. Deforce ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Bovine Serum Albumin ◽  
Horseradish Peroxidase ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Simultaneous Detection ◽  
Cross Reactivity ◽  
Efficient Approach ◽  
Selection For ◽  
Similar Affinity

Hapten heterology was introduced into the steps of hybridoma selection for the development of monoclonal antibodies (MAbs) against deoxynivalenol (DON). Firstly, a novel heterologous DON hapten was synthesised and covalently coupled to proteins (i.e. bovine serum albumin (BSA), ovalbumin and horseradish peroxidase) using the linkage of cyanuric chloride (CC). After immunisation, antisera from different DON immunogens were checked for the presence of useful antibodies. Next, both homologous and heterologous enzyme-linked immunosorbent assays were conducted to screen for hybridomas. It was found that heterologous screening could significantly reduce the proportion of false positives and appeared to be an efficient approach for selecting hybridomas of interest. This strategy resulted in two kinds of broad-selective MAbs against DON and its analogues. They were quite distinct from other reported DON-antibodies in their cross-reactivity profiles. A unique MAb 13H1 derived from DON-CC-BSA immunogen could recognise DON and its analogues in the order of HT-2 toxin ≯ 15-acetyl-DON ≯ DON ≯ nivalenol, with IC50 ranging from 1.14 to 7.69 μg/ml. Another preferable MAb 10H10 generated from DON-BSA immunogen manifested relatively similar affinity to DON, 3-acetyl-DON and 15-acetyl-DON, with IC50 values of 22, 15 and 34 ng/ml, respectively. This is the first broad-specific MAb against DON and its two acetylated forms and thus it can be used for simultaneous detection of the three mycotoxins.

scholarly journals Monoclonal antibodies to a phosphoprotein from chromatin of rat liver

1985 ◽  
Vol 225 (2) ◽  
pp. 357-363 ◽  
Author(s):  
M J Halikowski ◽  
C C Liew
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Bovine Serum Albumin ◽  
Rat Liver ◽  
High Specificity ◽  
Cross Reactivity ◽  
Dot Blot ◽  
Chromosomal Proteins ◽  
Rat Liver Nuclei ◽  
Liver Nuclei

Three monoclonal antibody subclasses (IgG1, IgG2a, and IgM) were raised to the phosphoprotein B2 (Mr 68000, pI6.5-8.2) which has been shown previously to be associated with the nucleosomes of rat liver nuclei. These antibodies do not show any significant cross reactivity with CM-cellulose ‘unbound’ non-histone chromosomal proteins, bovine serum albumin or histones. Further verification of the specificity of these antibodies to this phosphoprotein was carried out using both ‘dot’ blot and immunological transfer analysis (‘Western blot‘). The monoclonal antibodies (IgG1 and IgG2a) could also be used to semi-quantify the phosphoprotein B2 in rat liver nuclei. The high specificity and unlimited availability of this type of probe provides a means to study the role(s) of this phosphoprotein in the overall scheme of actively transcribed chromatin.

Monoclonal Antibodies and an Indirect ELISA for Detection of Psychrotrophic Bacteria in Refrigerated Milk

1997 ◽  
Vol 60 (1) ◽  
pp. 23-27 ◽  
Author(s):  
ROSALBA GUTIÉRREZ ◽  
ISABEL GONZÁLEZ ◽  
TERESA GARCÍA ◽  
ESTER CARRERA ◽  
BERNABÉ SANZ ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Horseradish Peroxidase ◽  
Indirect Elisa ◽  
Detection Threshold ◽  
Microtiter Plate ◽  
Live Cells ◽  
Elisa Assay ◽  
Psychrotrophic Bacteria ◽  
Milk Samples ◽  
Elisa Format

Monoclonal antibodies generated against live cells of Pseudomonas fluorescens have been used in an indirect ELISA format for the detection of Pseudomonas spp. and related psychrotrophic bacteria in refrigerated milk. The immunorecognition of monoclonal antibodies adsorbed to bacteria bound to the wells of a microtiter plate was performed with rabbit anti-mouse immunoglobulins conjugated to horseradish peroxidase. Subsequent enzymic conversion of the substrate resulted in distinct absorbance differences when assaying milk samples containing psychrotrophic bacteria in the range 105 to 109 CFU ml−1. The detection threshold for the ELISA assay developed in this work is 105 CFU ml−1.

scholarly journals Monoclonal antibody detection of Pseudomonas spp. in refrigerated meat by an indirect ELISA

1997 ◽  
Vol 24 (1) ◽  
pp. 5-8 ◽  
Author(s):  
R. Gutierrez ◽  
T. Garcia ◽  
I. González ◽  
B. Sanz ◽  
P. E. Hernández ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Indirect Elisa ◽  
Antibody Detection ◽  
Pseudomonas Spp

Generation of Human Monoclonal Anti-idiotypic Antibodies with Specificity to the Murine Monoclonal Anti-CA 125 Antibody B43.13

1996 ◽  
Vol 11 (4) ◽  
pp. 211-215
Author(s):  
J.B. Oltrogge ◽  
B. Donnerstag ◽  
R.P. Baum ◽  
A.A. Noujaim ◽  
L. Träger
Keyword(s):  
Ovarian Cancer ◽  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Peripheral Blood Lymphocytes ◽  
Cross Reactivity ◽  
Tumor Burden ◽  
Ca 125 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Monoclonal Antibodies ◽  
Ebv Transformation

Two human monoclonal antibodies, HID-7E7 and ROB-6F2, were produced by EBV transformation of peripheral blood lymphocytes (PBL). PBL were obtained from a patient with ovarian cancer who had been exposed several times to a Tc-99m labeled murine monoclonal anti-CA 125 antibody (B43.13, Biomira, Edmonton) for immunoscintigraphy. The HID-7E7 and ROB-6F2 producing B-cells were cloned with a limiting dilution technique and have shown stable immunoglobulin secretion within a period of three years. The human monoclonal antibodies HID-7E7 and ROB-6F2 are of the IgG isotype, and bind with significant affinity to the murine monoclonal antibody B43.13, which was used for immunoscintigraphy. Binding affinity of ROB-6F2 to other murine antibodies could not be detected. Cross reactivity of HID-7E7 to a murine anti-CEA monoclonal antibody was observed. In order to verify the anti-idiotypic character of the generated human antibodies, the ability of HID-7E7 and ROB-6F2, respectively, to inhibit the formation of the CA125/B43.13 complex is demonstrated via an enzyme-linked immunosorbent assay. These human anti-idiotypic antibodies are possible candidates for immunotherapy of ovarian cancer in patients with a small tumor burden following surgery and/or chemotherapy.

Development of antibodies for N-(1-deoxy-D-fructos-1-yl) fumonisin B1 and cross-reaction with modified fumonisins

2018 ◽  
Vol 11 (4) ◽  
pp. 493-502 ◽  
Author(s):  
C.M. Maragos ◽  
K.K. Sieve ◽  
M. Busman
Keyword(s):  
Monoclonal Antibodies ◽  
Indirect Elisa ◽  
Fumonisin B1 ◽  
Cross Reactivity ◽  
Cross Reaction ◽  
Immunoaffinity Column ◽  
Carbon Backbone ◽  
Matrix Components ◽  
Covalent Modifications

Fumonisins are a group of mycotoxins that are routinely found worldwide in commodities such as maize. The group, which has many members, is generally characterised by the presence of one or more tricarballylic acid groups esterified to a long carbon backbone. The diversity of this group of toxins is further augmented by their ability to interact with matrix components non-covalently and to form covalent products with matrix constituents, such as carbohydrates and proteins. Covalent modifications to the toxins make it more difficult to assess the total amounts that may be present in a commodity. We developed monoclonal antibodies (Mabs) against a known product of the reaction of fumonisin B1 (FB1) with glucose: N-(1-deoxy-D-fructos-1-yl) fumonisin B1 (NDFrc-FB1). Similar reactions were used to produce fructosyl-analogs of fumonisins B2 and B3, as well as galactose, maltose, and rhamnose analogs of FB1. These analogs were tested in a competitive indirect ELISA for cross-reactivity towards one of the developed antibodies (Mab 213221). All of the carbohydrate analogs cross-reacted with the Mab, at levels ranging from 75% (the FB3 analog derived from D-glucose) to 181% (the FB1 analog derived from maltose). These results suggested the assay was capable of binding to a wide variety of fumonisin-carbohydrate derivatives. The same antibody was incorporated into an immunoaffinity column that was used to isolate modified fumonisins from a sample of naturally contaminated maize. These results demonstrate the potential to isolate and detect modified fumonisins and will facilitate efforts to determine the frequency of the occurrence of these compounds in maize.

Production and Characterization of Antibodies Against Neosaxitoxin

1992 ◽  
Vol 75 (2) ◽  
pp. 341-345 ◽  
Author(s):  
Fun S Chu ◽  
Xuan Huang ◽  
Sherwood Hall
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Indirect Elisa ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Antibody ◽  
Antibody Titers ◽  
Antibody Elisa

Abstract Antibody against neosaxitoxin (neo-STX) was obtained from rabbits after immunization with neo- STX conjugated to either keyhole limpet hemocyanln (KLH) or bovine serum albumin (BSA). An indirect enzyme-linked Immunosorbent assay (ELISA), In which either neo-STX-BSA or neo-STXKLH was coated to the mlcroplate, was used to monitor the antibody titer. Although high antibody titers were obtained from rabbits after immunization with both immunogens, only antibody obtained from rabbits immunized with neo-STX-KLH was useful for Immunoassay. Competitive indirect ELISA revealed that the antibodies obtained from rabbits Immunized with neo-STX-KLH are specific for neo-STX but also have good cross-reactivity with STX. The concentrations causing 50% inhibition binding of neo-STX-BSA to the anti-neo-KLH by neo-STX, STX, and decarbamoyl-STX (DC-STX) were 0.9,8.0, and 53.1 ng/mL, respectively. Saxitoxin conjugated to polylyslne (STX-PLL) was also used as the coating reagent In the indirect ELISA. The concentrations causing 50% inhibition binding of antl-neo-STX-KLH to STX-PLL coated on the mlcrotiter plate by neo-STX, STX, and DC-STX were 1.2,4.1, and 36.1 ng/mL, respectively. With this newly developed antibody, ELISA could be a very effective method for monitoring seafood for both neo-STX and STX.

scholarly journals Development of Aflatoxin B1-Lysine Adduct Monoclonal Antibody for Human Exposure Studies

2001 ◽  
Vol 67 (6) ◽  
pp. 2712-2717 ◽  
Author(s):  
Jia-Sheng Wang ◽  
Salahaddin Abubaker ◽  
Xia He ◽  
Guiju Sun ◽  
Paul T. Strickland ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
High Risk ◽  
Bovine Serum Albumin ◽  
Dna Adducts ◽  
Human Exposure ◽  
Limit Of Detection ◽  
Serum Samples ◽  
Bovine Serum Albumin Conjugate ◽  
Aflatoxin B

ABSTRACT Mouse monoclonal antibodies were developed against a synthetic aflatoxin B1 (AFB)-lysine–cationized bovine serum albumin conjugate. The isotype of one of these antibodies, IIA4B3, has been classified as immunoglobulin G1(λ). The affinity and specificity of IIA4B3 were further characterized by a competitive radioimmunoassay. The affinities of IIA4B3 for AFB and its associated adducts and metabolites are ranked as follows: AFB-lysine > 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-AFB > AFB = 8,9-dihydro-8-(N 7-guanyl)-9-hydroxy-AFB > aflatoxin M1 > aflatoxin Q1. IIA4B3 had about a 10-fold higher affinity for binding to AFB-lysine adduct than to AFB when 3H-AFB–lysine was used as the tracer. The concentration for 50% inhibition for AFB-lysine was 0.610 pmol; that for AFB was 6.85 pmol. IIA4B3 had affinities at least sevenfold and twofold higher than those of 2B11, a previously developed antibody against parent AFB, for the major aflatoxin-DNA adducts 8,9-dihydro-8-(N 7-guanyl)-9-hydroxy-AFB and 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-AFB, respectively. An analytical method based on a competitive radioimmunoassay with IIA4B3 and3H-AFB–lysine was validated with a limit of detection of 10 fmol of AFB-lysine adduct. The method has been applied to the measurement of AFB-albumin adduct levels in human serum samples collected from the residents of areas at high risk for liver cancer.

scholarly journals Preparation and use of monoclonal antibodies  to detect Phytophthora nicotianae  var. nicotianae

1999 ◽  
Vol 35 (No. 2) ◽  
pp. 41-46
Author(s):  
B. Pekárová-Kyněrová ◽  
M. Kutíková
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Fusarium Oxysporum ◽  
Indirect Elisa ◽  
Phytophthora Nicotianae ◽  
Clavibacter Michiganensis ◽  
Tomato Plants ◽  
Clavibacter Michiganensis Subsp

A monoclonal antibody (MAb 18) was prepared against purified mycelial proteins from Phytophthora nicotianae var. nicotianae. The specificity of MAb 18 (lgG class) was tested using indirect ELISA (PTA-ELISA).It cross-reacted with Phytophthora cacto­ rum, P. cinrzamomi, P. cryptogea, P. fragariae) but not with other fungi (Fusarium  oxysporum, Pythium ultimwn and P. oligan­ drwn) and bacteria (Clavibacter michiganensis subsp. michiganensis) isolated from tomato. Phytophthora nicotianae var. nicotianae was detected in roots and basal stems of artificially infected young tomato plants using indirect ELISA and immunoprinting.

scholarly journals Anti-Lipid A Monoclonal Antibody Centoxin (HA-1A) Binds to a Wide Variety of Hydrophobic Ligands

1998 ◽  
Vol 66 (2) ◽  
pp. 870-873 ◽  
Author(s):  
E. J. Helmerhorst ◽  
J. J. Maaskant ◽  
B. J. Appelmelk
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Bovine Serum Albumin ◽  
Gel Electrophoresis ◽  
Lipid A ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Binding Properties ◽  
Antibody Epitopes

ABSTRACT This note describes the binding specificities of four lipid A monoclonal antibodies (MAbs) including Centoxin (HA-1A); these MAbs display similar binding properties. MAbs reacted with lipid A and heat-killed smooth bacteria, whereas no reactivity was observed with smooth lipopolysaccharide (LPS). Immunoblotting of bacterial extracts separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the MAbs bound to many polypeptide bands including the molecular weight markers. Denaturation of bovine serum albumin (BSA) by boiling or dithiothreitol treatment unmasked antibody epitopes. In addition, binding both to a hydrophobic aliphatic C12 chain covalently coupled to BSA and to single-stranded DNA was observed. The polyreactivity of these clones is most likely mediated by a preferential reactivity with hydrophobic molecular patches.

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