Evidence of Cross-Reactivity of ‘Carcinoma-Specific’ KC4 Monoclonal Antibody with Activated Mesothelial Cells and Phytohemagglutinin-Stimulated Lymphocytes

Oncology ◽  
1988 ◽  
Vol 45 (5) ◽  
pp. 380-383
Author(s):  
A. Neubauer ◽  
R. Musch ◽  
U. Thalmann ◽  
H. Grosser ◽  
J. Laser ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cross Reactivity ◽  
Mesothelial Cells

scholarly journals Generation of a Monoclonal Antibody against D-Dimer Using HTS-Based LiCA

2019 ◽  
Vol 25 (3) ◽  
pp. 310-319
Author(s):  
Yuan Dong ◽  
Hanjin Hou ◽  
An Chen ◽  
Wei Ma ◽  
Moli Yin ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Degradation Products ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cross Reactivity ◽  
Enzyme Hydrolysis ◽  
Clinical Samples ◽  
Sds Page ◽  
Thrombotic Diseases ◽  
D Dimer

D-dimer is an essential diagnostic index of thrombotic diseases. Since the existing anti-D-dimer antibodies vary in quality and specificity, a search for alternative anti-D-dimer antibodies is required. The present study aimed to screen a novel monoclonal antibody (mAb) against D-dimer using a light-initiated chemiluminescence assay (LiCA). In this work, mice were immunized with antigen prepared from human plasma by enzyme hydrolysis. After screening, a novel mAb, DD 2G11, was obtained. The results of sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis indicated that DD 2G11 could be used as a standard marker for D-dimer. The isotype of DD 2G11 was IgG1, the Ka value was 0.646 nM-1, and the Kd value was 50 nM, indicating that the binding affinity to D-dimer was very high. Furthermore, no cross-reactivity between DD 2G11 and other fibrinogen degradation products (FgDPs) was found. Finally, the correlation between DD 2G11 and the reference antibody (commercial antibody) was investigated by analyzing 56 clinical samples using a latex-enhanced turbidimetric immunoassay (LTIA). The R2 value of the linear regression was 0.94538, indicating that DD 2G11 met clinical requirements. In conclusion, the present study provides a more expeditious protocol to screen mAbs and provides a clinically usable mAb against D-dimer.

A validation study of selected methods routinely used for measurement of cyclosporine

1990 ◽  
Vol 36 (4) ◽  
pp. 670-674 ◽  
Author(s):  
K E Blick ◽  
S H Melouk ◽  
H D Fry ◽  
R L Gillum
Keyword(s):  
Monoclonal Antibody ◽  
Polyclonal Antibody ◽  
Validation Study ◽  
Liver Dysfunction ◽  
Correlation Coefficients ◽  
Cross Reactivity ◽  
Poor Correlation ◽  
Fluorescence Polarization Immunoassay ◽  
Gamma Glutamyltransferase ◽  
Transplant Patients

Abstract We compare four methods for measuring cyclosporine (CyA) in plasma and whole blood of transplant patients: HPLC, RIA with a polyclonal antibody, RIA with a monoclonal antibody, and fluorescence polarization immunoassay (FPIA). The monoclonal RIA procedure correlated acceptably with HPLC, with slope = 1.21, r = 0.97, and Sy,x = +/- 40.1. However, the FPIA, done in three separate instruments, correlated relatively poorly with HPLC, giving slopes of 1.67, 1.51, and 2.32; correlation coefficients of 0.72, 0.43, and 0.83; and Sy,x = +/- 205.4, +/- 334.5, and +/- 222.4. The polyclonal RIA correlated reasonably well with HPLC, with a slope = 1.15, r = 0.90, and Sy,x = +/- 72.6. Values for individual patients with increases both in gamma-glutamyltransferase and creatinine showed very poor correlation between FPIA and HPLC, which suggests that metabolite cross-reactivity with FPIA is significant and unpredictable in patients with liver dysfunction coexisting with renal dysfunction. Evidently, the monoclonal RIA can be substituted for HPLC, if the therapeutic range is adjusted for the 21% higher results obtained by RIA.

scholarly journals Structure-Based Modification of an Anti-neuraminidase Human Antibody Restores Protection Efficacy against the Drifted Influenza Virus

mBio ◽  
2020 ◽  
Vol 11 (5) ◽  
Author(s):  
Haihai Jiang ◽  
Weiyu Peng ◽  
Jianxun Qi ◽  
Yan Chai ◽  
Hao Song ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Influenza Virus ◽  
Antibody Fragment ◽  
Salt Bridge ◽  
Cross Reactivity ◽  
Human Antibody ◽  
Influenza Strain ◽  
Group 2 ◽  
Fragment Antigen Binding ◽  
Group 1

ABSTRACT Here, we investigate a monoclonal antibody, Z2B3, isolated from an H7N9-infected patient, that exhibited cross-reactivity to both N9 (group 2) and a broad range of seasonal and avian N1 (group 1) proteins but lost activity to the N1 with the substitution K432E. This substitution exists in 99.25% of seasonal influenza strains after 2013. The NA-Z2B3 complex structures indicated that Z2B3 binds within the conserved active site of the neuraminidase (NA) protein. A salt bridge between D102 in Z2B3 and K432 in NA plays an important role in binding. Structure-based modification of Z2B3 with D102R in heavy chain reversed the salt bridge and restored the binding and inhibition of N1 with E432. Furthermore, Z2B3-D102R can protect mice from A/Serbia/NS-601/2014 H1N1 virus (NA contains E432) infection while the wild-type Z2B3 antibody shows no protection. This study demonstrates that a broadly reactive and protective antibody to NA can be in principle edited to restore binding and inhibition to recently drifted N1 NA and regain protection against the variant influenza strain. IMPORTANCE The immune system produces antibodies to protect the human body from harmful invaders. The monoclonal antibody (MAb) is one kind of effective antivirals. In this study, we isolated an antibody (Z2B3) from an H7N9 influenza virus-infected child. It shows cross-reactivity to both group 1 (N1) and group 2 (N9) neuraminidases (NAs) but is sensitive to N1 NA with a K432E substitution. Structural analysis of the NA-antibody fragment antigen-binding (Fab) complex provides a clue for antibody modification, and the modified antibody restored binding and inhibition to recently drifted N1 NA and regained protection against the variant influenza strain. This finding suggests that antibodies to NA may be a useful therapy and can be in principle edited to defeat drifted influenza virus.

Listeriolysin O-Based Latex Agglutination Test for the Rapid Detection of Listeria monocytogenes in Foods

1997 ◽  
Vol 60 (9) ◽  
pp. 1038-1040 ◽  
Author(s):  
GHASSAN M. MATAR ◽  
PEGGY S. HAYES ◽  
WILLIAM F. BIBB ◽  
BALA SWAMINATHAN
Keyword(s):  
Monoclonal Antibody ◽  
Listeria Monocytogenes ◽  
Rapid Detection ◽  
Rapid Test ◽  
Food Samples ◽  
Cross Reactivity ◽  
Latex Agglutination ◽  
Listeriolysin O ◽  
Agglutination Assay ◽  
Latex Beads

A latex agglutination-based test for the rapid detection of Listeria monocytogenes in foods was developed. An antilisteriolysin O (LLO) monoclonal antibody (HID5E12D7; IgG2b) covalently bound to polystyrene amidine-modified latex beads was used in a slide agglutination assay. The latex reagent detected 0.1 ng/ml of LLO in phosphate-buffered saline plus bovine serum albumin. It reacted with culture supernatants of L. monocytogenes but not with other Listeria species or Streptococcus groups A through G. The listeriolysin O latex agglutination assay (LLOLAT) was applied to 24-h and 48-h USDA primary enrichment cultures of 208 food samples obtained from refrigerators of listeriosis patients enrolled in a study to determine the role of foods in sporadic listeriosis. Of 19 samples positive by cultural techniques, 17 were positive by the LLOLAT. Cultures with low (<0.3 CFU/g) levels of L. monocytogenes were positive in the LLOLAT. No cross-reactivity occurred when using a heterogeneous monoclonal antibody. The LLOLAT is a sensitive, specific and rapid test and may be useful for screening foods for L. monocytogenes.

scholarly journals A NS1-binding monoclonal antibody interacts with two residues that are highly conserved in seasonal as well as newly emerged influenza A virus

2019 ◽  
Vol 77 (1) ◽  
Author(s):  
Su Hui Catherine Teo ◽  
Jian-Ping Wu ◽  
Chee-Keng Mok ◽  
Yee-Joo Tan
Keyword(s):  
Monoclonal Antibody ◽  
Influenza A Virus ◽  
Influenza A ◽  
Rna Binding ◽  
Intracellular Localization ◽  
Cross Reactivity ◽  
Structural Protein ◽  
Multifunctional Protein ◽  
Good Target ◽  
Conformational Plasticity

Abstract The non-structural protein 1 (NS1) of influenza A virus (IAV) is a multifunctional protein that antagonizes host antiviral responses, modulating virus pathogenesis. As such, it serves as a good target for research and diagnostic assay development. In this study, we have generated a novel monoclonal antibody (mAb) 19H9 and epitope mapping revealed that two residues, P85 and Y89, of NS1 are essential for interacting with this mAb. Furthermore, residues P85 and Y89 are found to be highly conserved across different IAV subtypes, namely seasonal H1N1 and H3N2, as well as the highly pathogenic H5N1 and H5N6 avian strains. Indeed, mAb 19H9 exhibits broad cross-reactivity with IAV strains of different subtypes. The binding of mAb 19H9 to residue Y89 was further confirmed by the abrogation of interaction between NS1 and p85β. Additionally, mAb 19H9 also detected NS1 proteins expressed in IAV-infected cells, showing NS1 intracellular localization in the cytoplasm and nucleolus. To our knowledge, mAb 19H9 is the first murine mAb to bind at the juxtaposition between the N-terminal RNA-binding domain and C-terminal effector domain of NS1. It could serve as a useful research tool for studying the conformational plasticity and dynamic changes in NS1.

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.

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.

Amplified Enzyme-Linked Immunoassay of Human Proinsulin in Serum (Detection Limit: 0.1 pmol/L)

1992 ◽  
Vol 38 (2) ◽  
pp. 227-232 ◽  
Author(s):  
F J Dhahir ◽  
D B Cook ◽  
C H Self
Keyword(s):  
Monoclonal Antibody ◽  
Alkaline Phosphatase ◽  
Cross Reactivity ◽  
C Peptide ◽  
Human Proinsulin ◽  
Microtiter Plates ◽  
Enzyme Amplification ◽  
Fold Excess ◽  
Amplification Procedure ◽  
Serum Detection

Abstract We describe an amplified enzyme-linked immunoassay of human proinsulin in serum that detects intact proinsulin and both the 32/33 and 65/66 split forms. The method uses the IgG fraction of a polyclonal antibody raised in a guinea pig against intact proinsulin, which we used to coat plastic microtiter plates. A sandwich was formed with proinsulin by using a monoclonal antibody against C-peptide labeled with alkaline phosphatase. We quantified the reaction by using the enzyme amplification procedure, which detected as little intact proinsulin as 0.1 pmol/L. We found no cross-reactivity with C-peptide in the assay, and decreased recovery attributable to the presence of insulin could be demonstrated only with a 30-fold excess of this hormone over proinsulin.

scholarly journals Development of Human Monoclonal Antibody for Claudin-3 Overexpressing Carcinoma Targeting

Biomolecules ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 51 ◽  
Author(s):  
Hobin Yang ◽  
Hayeon Park ◽  
Yong Jin Lee ◽  
Jun Young Choi ◽  
TaeEun Kim ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Transmembrane Domain ◽  
Malignant Tumors ◽  
Human Monoclonal Antibody ◽  
Cross Reactivity ◽  
Drug Conjugates ◽  
Igg1 Monoclonal Antibody ◽  
Potential Biomarker ◽  
Human Igg1 ◽  
Mediate Cell

Most malignant tumors originate from epithelial tissues in which tight junctions mediate cell–cell interactions. Tight junction proteins, especially claudin-3 (CLDN3), are overexpressed in various cancers. Claudin-3 is exposed externally during tumorigenesis making it a potential biomarker and therapeutic target. However, the development of antibodies against specific CLDN proteins is difficult, because CLDNs are four-transmembrane domain proteins with high homology among CLDN family members and species. Here, we developed a human IgG1 monoclonal antibody (h4G3) against CLDN3 through scFv phage display using CLDN3-overexpressing stable cells and CLDN3-embedded lipoparticles as antigens. The h4G3 recognized the native conformation of human and mouse CLDN3 without cross-reactivity to other CLDNs. The binding kinetics of h4G3 demonstrated a sub-nanomolar affinity for CLDN3 expressed on the cell surface. The h4G3 showed antibody-dependent cellular cytotoxicity (ADCC) according to CLDN3 expression levels in various cancer cells by the activation of FcγRIIIa (CD16a). The biodistribution of h4G3 was analyzed by intravenous injection of fluorescence-conjugated h4G3 which showed that it localized to the tumor site in xenograft mice bearing CLDN3-expressing tumors. These results indicate that h4G3 recognizes CLDN3 specifically, suggesting its value for cancer diagnosis, antibody-drug conjugates, and potentially as a chimeric antigen receptor (CAR) for CLDN3-expressing pan-carcinoma.

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