Production and Characterization of a New Monoclonal Antibody AgainstNeisseria meningitidis: Study of the Cross-Reactivity with Different Bacterial Genera

Hybridoma ◽  
2000 ◽  
Vol 19 (6) ◽  
pp. 445-453 ◽  
Author(s):  
Elizabeth Natal De Gaspari
Keyword(s):  
Monoclonal Antibody ◽  
Cross Reactivity ◽  
The Cross

Characterization of G12 Sandwich ELISA, a Next-Generation Immunoassay for Gluten Toxicity

2012 ◽  
Vol 95 (2) ◽  
pp. 372-376 ◽  
Author(s):  
Elisabeth Halbmayr-Jech ◽  
Elisabeth Hammer ◽  
Richard Fielder ◽  
Jacqueline Coutts ◽  
Adrian Rogers ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Celiac Disease ◽  
Working Group ◽  
Sandwich Elisa ◽  
Extraction Methods ◽  
Cross Reactivity ◽  
Next Generation ◽  
Preliminary Results ◽  
Sample Extraction

Abstract In this work, a monoclonal antibody called G12, raised against the most immunotoxic peptide to celiac disease patients, was used to develop a sandwich ELISA. Preliminary results on cross-reactivities, recoveries, and extraction methods of the new assay are presented. The assay calibration was performed using material from the Prolamin Working Group. The antibody's specificity was determined by cross-reactivity studies on different grains, nuts, oils, and starches. Recovery of the assay was determined by spiking experiments on common food matrixes, as well as on problematic matrixes. Furthermore, sample extraction methods using ethanol, cocktail solution, and a proprietary buffer have been compared.

IgE response in schistosomiasis japonica: characterization of a cercarial allergen in comparison with purified egg allergens

1997 ◽  
Vol 71 (3) ◽  
pp. 221-226 ◽  
Author(s):  
M. Owhashi ◽  
Y. Horii ◽  
A. Ishii
Keyword(s):  
Molecular Weight ◽  
Apparent Molecular Weight ◽  
Cross Reactivity ◽  
Schistosomiasis Japonica ◽  
Gel Chromatography ◽  
Con A ◽  
The Cross ◽  
Ige Response ◽  
The Relationship

AbstractThe relationship between the cercarial allergen and two previously isolated egg allergens (J1, J2) of Schistosoma japonicum was examined especially in terms of the cross-reactivity between them. Semi-purified cercarial allergen (JAC) was obtained from the crude extract of S. japonicum cercariae by gel chromatography on Sephadex G-200. The apparent molecular weight of JAC was estimated approximately as 60–100 kDa. JAC could bind to Con A-Sepharose, indicating its glycoprotein nature. Three groups of BALB/c mice were immunized with JAC, J1 or J2 using A1(OH)3 as adjuvant, and the cross-reactivity of each antiserum was examined by PCA. Anti-JAC, anti-Jl or anti-J2 serum was highly specific to the corresponding antigen. When IgE-ELISA of S. japonicum patient sera was performed using JAC, J1 or J2 as an antigen, the correlation between anti-J1 and anti-J2 (r = 0.78) was high, whereas the correlation between anti-JAC and anti-J1 (r = 0.27) or between anti-JAC and anti-J2 (r = 0.12) was low. These results suggest that most IgE epitopes on cercarial allergen are independent from those on egg allergens in S. japonicum.

Characterization of a monoclonal antibody (HB-22) and development of an ELISA for human apolipoprotein A-I

1995 ◽  
Vol 41 (8) ◽  
pp. 1150-1158 ◽  
Author(s):  
T C Rothwell ◽  
V S Kamanna ◽  
F Y Jin ◽  
E Koren ◽  
T Foley ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Binding Capacity ◽  
Cross Reactivity ◽  
High Density Lipoproteins ◽  
Tween 20 ◽  
Major Protein ◽  
Binding Studies ◽  
Very Low Density Lipoproteins ◽  
Human Apolipoprotein

Abstract We describe the production and characterization of a high-affinity monoclonal antibody, HB-22, for apolipoprotein (apo) A-I, a major protein of human high-density lipoproteins (HDL). Including Tween 20 in the reaction mixture increased the binding capacity of HB-22 to apo A-I. HB-22 showed monospecific reactivity with HDL or apo A-I, displaying no cross-reactivity with apo A-II, intermediate-, low-, or very-low-density lipoproteins. Immunoaffinity columns with HB-22 (in the absence of Tween 20) showed an immunosorbent capacity of 80 micrograms of apo A-I per milligram of antibody. The immunosorbent capacity of HB-22 for apo A-I was similar in plasma samples from normolipidemic, hypercholesterolemic, or hypertriglyceridemic patients. Comparative binding studies demonstrated that compared with other available monoclonal apo A-I antibodies, HB-22 had the greatest apparent affinity for binding to HDL. A competitive ELISA developed by utilizing HB-22 could detect as little as 20 ng of apo A-I in the reaction mixture. The intra- and interassay CVs of the ELISA were 5.4% and 9.5%, respectively.

scholarly journals Potent binding of 2019 novel coronavirus spike protein by a SARS coronavirus-specific human monoclonal antibody

2020 ◽  
Author(s):  
Xiaolong Tian ◽  
Cheng Li ◽  
Ailing Huang ◽  
Shuai Xia ◽  
Sicong Lu ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Binding Site ◽  
Neutralizing Antibodies ◽  
Rapid Development ◽  
Antiviral Treatment ◽  
Human Monoclonal Antibody ◽  
Cross Reactivity ◽  
Spike Protein ◽  
The Cross ◽  
Novel Coronavirus

ABSTRACTThe newly identified 2019 novel coronavirus (2019-nCoV) has caused more than 800 laboratory-confirmed human infections, including 25 deaths, posing a serious threat to human health. Currently, however, there is no specific antiviral treatment or vaccine. Considering the relatively high identity of receptor binding domain (RBD) in 2019-nCoV and SARS-CoV, it is urgent to assess the cross-reactivity of anti-SARS-CoV antibodies with 2019-nCoV spike protein, which could have important implications for rapid development of vaccines and therapeutic antibodies against 2019-nCoV. Here, we report for the first time that a SARS-CoV-specific human monoclonal antibody, CR3022, could bind potently with 2019-nCoV RBD (KD of 6.3 nM). The epitope of CR3022 does not overlap with the ACE2 binding site within 2019-nCoV RBD. Therefore, CR3022 has the potential to be developed as candidate therapeutics, alone or in combination with other neutralizing antibodies, for the prevention and treatment of 2019-nCoV infections. Interestingly, some of the most potent SARS-CoV-specific neutralizing antibodies (e.g., m396, CR3014) that target the ACE2 binding site of SARS-CoV failed to bind 2019-nCoV spike protein, indicating that the difference in the RBD of SARS-CoV and 2019-nCoV has a critical impact for the cross-reactivity of neutralizing antibodies, and that it is still necessary to develop novel monoclonal antibodies that could bind specifically to 2019-nCoV RBD.

scholarly journals Production and Characterization of Monoclonal Antibodies against the Hemolysin BL Enterotoxin Complex Produced byBacillus cereus

1999 ◽  
Vol 65 (10) ◽  
pp. 4470-4474 ◽  
Author(s):  
R. Dietrich ◽  
C. Fella ◽  
S. Strich ◽  
E. Märtlbauer
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Bacillus Cereus ◽  
Cell Lines ◽  
Cross Reactivity ◽  
Uncharacterized Protein ◽  
Hybridoma Cell Lines ◽  
Different Strains ◽  
Hemolysin Bl

ABSTRACT A total of five hybridoma cell lines that produced monoclonal antibodies against the components of the hemolysin BL (HBL) enterotoxin complex and sphingomyelinase produced by Bacillus cereuswere established and characterized. Monoclonal antibody 2A3 was specific for the B component, antibodies 1A12 and 8B12 were specific for the L2 component, and antibody 1C2 was specific for the L1 protein of the HBL enterotoxin complex. No cross-reactivity with other proteins produced by different strains ofB. cereus was observed for monoclonal antibodies 2A3, 1A12, and 8B12, whereas antibody 1C2 cross-reacted with an uncharacterized protein of approximately 93 kDa and with a 39-kDa protein, which possibly represents one component of the nonhemolytic enterotoxin complex. Antibody 2A12 finally showed a distinct reactivity withB. cereus sphingomyelinase. The monoclonal antibodies developed in this study were also successfully applied in indirect enzyme immunoassays for the characterization of the enterotoxic activity of B. cereus strains. About 50% of the strains tested were capable of producing the HBL enterotoxin complex, and it could be demonstrated that all strains producing HBL were also highly cytotoxic.

Isolation and preliminary characterization of a monoclonal antibody that interacts preferentially with the liver isoenzyme of human alkaline phosphatase.

1985 ◽  
Vol 31 (3) ◽  
pp. 381-385 ◽  
Author(s):  
G M Lawson ◽  
J A Katzmann ◽  
T K Kimlinger ◽  
J F O'Brien
Keyword(s):  
Monoclonal Antibody ◽  
Alkaline Phosphatase ◽  
Human Liver ◽  
Clinical Laboratory ◽  
Bone Alkaline Phosphatase ◽  
Cross Reactivity ◽  
Immunological Methods ◽  
Enzymic Assay ◽  
Murine Monoclonal Antibodies

Abstract We have prepared murine monoclonal antibodies against isolated human bone alkaline phosphatase (ALP, EC 3.1.3.1). Hybridoma supernates were separately screened for reactivity against both human liver and bone ALP. Although most antibody-positive hybrids showed similar reactivity against both isoenzymes, one hybridoma produced an antibody that interacted preferentially with liver ALP. This antibody was purified and used to establish an immunoassay to differentiate liver ALP from bone ALP. When equal activities of the two isoenzymes (as determined by a conventional enzymic assay) were measured by the immunoassay, a fivefold greater response was obtained with liver than with bone ALP. The immunoassay can be used to measure the proportions of the bone and liver isoenzymes in mixtures of them. Cross reactivity with human placental and intestinal ALP is less than 3% relative to liver ALP. These findings support the feasibility of developing immunological methods to differentiate these isoenzymes in the clinical laboratory.

Characterization of Tumor Antigen in Epidermoid Carcinoma

1975 ◽  
Vol 84 (6) ◽  
pp. 787-792 ◽  
Author(s):  
Charles J. Krause ◽  
John Nysather ◽  
Brian F. McCabe
Keyword(s):  
Molecular Weight ◽  
Head And Neck ◽  
Isoelectric Focusing ◽  
Tumor Antigen ◽  
Tumor Antigens ◽  
Cross Reactivity ◽  
Epidermoid Carcinoma ◽  
The Cross

Tumor antigens have been isolated from nine epidermoid carcinomas of the head and neck. The most strongly reactive antigens have a molecular weight of ≃ 25,000–50,000 daltons, though other antigens weighing ≃ 120,000–400,000 daltons were noted in six of the tumors. The most strongly reactive antigens from three of the tumors were further purified by isoelectric focusing. Each of these antigens had a pI between 8.36 and 8.40. The cross reactivity of these antigens will be studied next. It is hoped that the purified tumor antigen will be useful in the development of an immunodetection system for epidermoid carcinoma.

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