Production and Characterization of Monoclonal Antibodies against Acridine Ester

2014 ◽  
Vol 1010-1012 ◽  
pp. 101-106
Author(s):  
Li Xin Zhu ◽  
Yan Fan ◽  
Xue Mei Qiu ◽  
Ren Rong Liu ◽  
Long Xu ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Keyhole Limpet Hemocyanin ◽  
Absorption Method ◽  
Weakly Alkaline ◽  
Indirect Competitive Elisa ◽  
Traditional Procedure ◽  
Inhibition Concentration ◽  
Alkaline Conditions

AE (Acridine ester) artificial antigens were obtained by acridine ester coupled with KLH (keyhole limpet hemocyanin) and BSA (bovine serum albumin) in weakly alkaline conditions, respectively. These two kinds of antigens were identified by the ultraviolet absorption method, which showed that they were synthesized successfully. Balb/c mice were immunized with the AE-KLH for producing the McAb (monoclonal antibody) against AE, and then cell fusion was used. Two monoclonal antibodies against AE were acquired according to traditional procedure. One of anti-AE antibodies produced from the 1E8 cell lines was the IgG3 subclass with a kappa-type light chain. An indirect competitive ELISA assay was developed. A linear relationship was observed over the concentration range from 0.25 to 10 ng/mL, and the detection limit was 0.125 ng/mL. The 50% inhibition concentration was 2.3 ng/mL. The monoclonal antibody against AE was successfully obtained.

scholarly journals Production and characterization of monoclonal antibodies to the edta extract of Leptospira interrogans, serovar icterohaemorrhagiae

1996 ◽  
Vol 29 (5) ◽  
pp. 483-489
Author(s):  
Lilian Terezinha de Queiroz Leite ◽  
Mauricio Resende ◽  
Wanderley de Souza ◽  
Elizabeth R.S. Camargos ◽  
Matilde Cota Koury
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Western Blot ◽  
Silver Staining ◽  
Colloidal Gold ◽  
Leptospira Interrogans ◽  
Outer Envelope ◽  
Lethal Challenge ◽  
Sds Page

Monoclonal antibodies (MABs) ivere produced against an etbylenediaminetetraacetate (EDTA) extract of Leptospira interrogans serovar icterohaemorrhagiae being characterized by gel precipitation as IgM and IgG (IgGl and IgG2b). The EDTA extract was detected as several bands by silver staining in SDS-PAGE. In the Western blot the bands around 20 KDa reacted with a monoclonal antibody, 47B4D6, and was oxidized by periodate and was not digested by pronase, suggesting that the determinant is of carbohydrate nature, lmmunocytochemistry, using colloidal gold labeling, showed that an EDTA extract determinant recognized by monoclonal antibody 47B4D6, is localized under the outer envelope of serovar icterohaemorrhagiae. Hoe AIAB raised against the EDTA extract was not able to protect hamsters from lethal challenge with virulent homologous leptospires.

scholarly journals Isolation of monoclonal antibodies specific for products of avian oncogene myb.

1984 ◽  
Vol 4 (12) ◽  
pp. 2843-2850 ◽  
Author(s):  
G I Evan ◽  
G K Lewis ◽  
J M Bishop
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Nuclear Protein ◽  
Gene Products ◽  
Viral Oncogenes ◽  
Myb Gene

We isolated a series of monoclonal antibodies which were raised against a bacterially expressed protein, bp37v-myb, and coded for by part of the avian v-myb gene. These monoclonal antibodies recognized a range of antigenic specificities on bp37v-myb, and this was reflected in their differing specificities for the gene products of the v-myb, c-myb, and E26 viral oncogenes. One monoclonal antibody recognized, in addition to the v-myb and c-myb gene products, a conserved nuclear protein found in all tested cells. We describe the characterization of these monoclonal antibodies.

Generation and Characterization of a Murine Monoclonal Antibody Specific for the Human T1-Cd5 Molecule

1987 ◽  
Vol 2 (3) ◽  
pp. 143-150 ◽  
Author(s):  
Federico Genzano ◽  
Ada Funaro ◽  
Massimo Alessio ◽  
Lucia B. De Monte ◽  
Graziella Bellone ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
T Cell ◽  
T Lymphocytes ◽  
Membrane Glycoprotein ◽  
Functional Features ◽  
Specific Membrane ◽  
Murine Monoclonal Antibodies ◽  
Selection Of

Murine monoclonal antibodies (MoAbs) have found widespread applications in the characterization of the molecular and functional features of lymphocyte differentiation antigens. The present paper summarizes the results of our work dealing with the production and selection of a murine MoAb recognizing a molecule expressed during the whole differentiative life of T lymphocytes. The MoAb CB01 resulted to be specific for an apparently unique epitope of the T-cell specific membrane glycoprotein T1-CD5.

Production and Characterization of Antibodies against Fumonisin B1

1996 ◽  
Vol 59 (9) ◽  
pp. 992-997 ◽  
Author(s):  
FENG-YIR YU ◽  
FUN S. CHU
Keyword(s):  
Detection Limit ◽  
Polyclonal Antibodies ◽  
Fumonisin B1 ◽  
Correlation Coefficients ◽  
Keyhole Limpet Hemocyanin ◽  
Hplc Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Indirect Competitive Elisa ◽  
Antibody Titers

Polyclonal antibodies against fumonisin B1 (FmB1) were produced in rabbits after immunizing the animals with either FmBl-keyhole limpet hemocyanin (KLH) or FmB1 bovine serum albumin (BSA). A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and an indirect competitive ELISA (idc-ELISA) were used for the characterization of the antibodies and for analysis of the toxin in corn samples. The antibody titers in the serum of rabbits immunized with FmBl-KLH were considerably higher than in those immunized with FmBl-BSA. The antibodies from the rabbits immunized with FmBl-KLH were further characterized. The concentrations causing 50% inhibition of binding of FmB1-horseradish peroxidase (HRP) to the antibodies by FmB1, FmB2 and FmB3 in the ELISA were found to be 0.45, 0.72, and 25 ng/ml, respectively. The detection limit of FmBl, based on 95% confidence at 5% of inhibition of binding of FmBl-HRP conjugate, in buffer of the dc-ELISA was found to be 0.05 ng/ml. In the presence of a matrix such as corn, the detection limit was less than 50 ppb. The overall analytical recoveries of FmBl (50 to 1,000 ng/g) added to the ground corn and then extracted with CH3CN/H2O (1/1, vol/vol) with cleanup and without cleanup in the dc ELISA were found to be 70.5 and 85.9%, respectively. A good correlation was found between the FmBl levels in 2 starch and 10 naturally contaminated corn samples analyzed by the dc-ELISA and the high-pressure liquid chromatography (HPLC) method. The correlation coefficients between ELISA and HPLC were found to be 0.955 (y [ELISA] = 1.3 1x [HPLC] + 77 ppb; P < 0.001) and 0.811 (y = 1.13x + 34 ppb; P < 0.01) for the sample without and with cleanup treatment, respectively.

scholarly journals Characterization of a Broadly Reactive Monoclonal Antibody against Norovirus Genogroups I and II: Recognition of a Novel Conformational Epitope

2007 ◽  
Vol 81 (22) ◽  
pp. 12298-12306 ◽  
Author(s):  
Tomoyuki Shiota ◽  
Michio Okame ◽  
Sayaka Takanashi ◽  
Pattara Khamrin ◽  
Makiko Takagi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Conformational Epitope ◽  
The Novel ◽  
Antigen Specificity ◽  
Virus Like Particle ◽  
The Family ◽  
Immunological Diagnosis ◽  
Further Development

ABSTRACT Norovirus, which belongs to the family Caliciviridae, is one of the major causes of nonbacterial acute gastroenteritis in the world. The main human noroviruses are of genogroup I (GI) and genogroup II (GII), which were subdivided further into at least 15 and 18 genotypes (GI/1 to GI/15 and GII/1 to GII/18), respectively. The development of immunological diagnosis for norovirus had been hindered by the antigen specificity of the polyclonal antibody. Therefore, several laboratories have produced broadly reactive monoclonal antibodies, which recognize the linear GI and GII cross-reactive epitopes or the conformational GI-specific epitope. In this study, we characterized the novel monoclonal antibody 14-1 (MAb14-1) for further development of the rapid immunochromatography test. Our results demonstrated that MAb14-1 could recognize 15 recombinant virus-like particles (GI/1, 4, 8, and 11 and GII/1 to 7 and 12 to 15) and showed weak affinity to the virus-like particle of GI/3. This recognition range is the broadest of the existing monoclonal antibodies. The epitope for MAb14-1 was identified by fragment, sequence, structural, and mutational analyses. Both terminal antigenic regions (amino acid positions 418 to 426 and 526 to 534) on the C-terminal P1 domain formed the conformational epitope and were in the proximity of the insertion region (positions 427 to 525). These regions contained six amino acids responsible for antigenicity that were conserved among genogroup(s), genus, and Caliciviridae. This epitope mapping explained the broad reactivity and different titers among GI and GII. To our knowledge, we are the first group to identify the GI and GII cross-reactive monoclonal antibody, which recognizes the novel conformational epitope. From these data, MAb14-1 could be used further to develop immunochromatography.

Monoclonal antibodies against plasma protease inhibitors: II. Production and characterization of 25 monoclonal antibodies against human α1-antitrypsin. Correlation between antigenic structure and functional sites

1984 ◽  
Vol 4 (2) ◽  
pp. 139-147 ◽  
Author(s):  
P. Hérion ◽  
D. Siberdt ◽  
M. Francotte ◽  
J. Urbain ◽  
A. Bollen
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Protease Inhibitors ◽  
Antigenic Structure ◽  
Light Chains ◽  
Enzyme Linked Immunosorbent Assay ◽  
Functional Sites ◽  
Heavy Chains ◽  
Low Levels

Twenty-five hybridomas secreting monoclonal antibodies against human α1-antitrypsim have been produced by the cell-fusion techmque (Kóhler and Milstein, 1976). All antibodies are specific for α1-antitrypsim and carry γ1-antitrypsim heavy chains and κ light chains. Inhibition experiments showed that these monoclonal antibodies define three independent antigenic regions on the α1-antitrypsim molecule; one of these domains appears to be involved in the interaction between α1-antitrypsim and trypsin. In addition, one monoclonal antibody, AATY39, was used to develop an enzyme-linked immunosorbent assay capable of detecting low levels of α1-antitrypsim in the range of 1 to 2 ng/ml.

Characterization of a Giardia-like microorganism causing false positive reactions in the detection of Cryptosporidium/Giardia from water samples

2012 ◽  
Vol 12 (5) ◽  
pp. 604-610
Author(s):  
Taketoshi Shimizu ◽  
Takuya Oda ◽  
Hiroyuki Ito ◽  
Shoji Uga
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Dna Sequence ◽  
Water Samples ◽  
False Positive ◽  
18S Rdna ◽  
Giardia Cyst ◽  
Morphological Aspects ◽  
Detection Assay

Giardia-like microorganisms were found as a result of a Cryptosporidium/Giardia detection assay on winter reservoir samples. The morphological aspects, DNA sequence of the microorganism, and specificity of immunoassay reagents were investigated. The microorganism was very similar to a Giardia cyst. The cell was oval shaped (11.0 μm in length, 7.7 μm in width), contained organelles and was stained bright apple-green by a fluorescent monoclonal antibody, although no axonemes or curved bristles were observed. Cross-reaction occurred with the anti-Cryptosporidium, but not the anti-Giardia, immunomagnetic antibody. The specificity of the fluorescent monoclonal antibodies differed depending on the product. As a result of the analysis of the partial sequence of 18S rDNA, the microorganism was identified as Lunulospora curvula, mitosporic Ascomycota. These findings will be very helpful for avoiding false positives in future assays.

Characterization of monoclonal antibodies directed against the TSH receptor, as revealed with the cytochemical bioassay

1987 ◽  
Vol 116 (1_Suppl) ◽  
pp. S173-S180 ◽  
Author(s):  
N.J. Marshall ◽  
L. D. Kohn ◽  
P. A. Ealey
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Tsh Receptor ◽  
Thyroid Cells ◽  
Essential Component ◽  
Potent Stimulator ◽  
Cytochemical Bioassay

Abstract. The monoclonal antibody 11E8 (Kohn et al. 1984) is a novel proof of the actions of thyroid stimulator, since it is not only a potent stimulator of thyroid cells but also an inhibitor for TSH-binding. This might reflect an interaction of 11E8 with a discrete domaine of the TSH receptor, which is an essential component for TSH but not for TSab actions. These unique properties of 11E8 can be applied to the characterization of other thyroid stimulators which may interact with different domaines of the receptor. Moreover, 'TSab-like' stimulators, such as the sheep anti-TSH antiserum, could potentially be identified by 11E8.

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