Characterization of the In Vitro and In Vivo Activity of Monoclonal Antibodies to Human IL-18

Hybridoma ◽  
2000 ◽  
Vol 19 (5) ◽  
pp. 363-367 ◽  
Author(s):  
Steve Holmes ◽  
Julie A. Abrahamson ◽  
Niam Al-Mahdi ◽  
Sherin S. Abdel-Meguid ◽  
Yen Sen Ho
Keyword(s):  
Monoclonal Antibodies

scholarly journals Generation and Characterization of Typhoid Toxin-Neutralizing Human Monoclonal Antibodies

2020 ◽  
Vol 88 (10) ◽  
Author(s):  
Xuyao Jiao ◽  
Sarah Smith ◽  
Gabrielle Stack ◽  
Qi Liang ◽  
Allan Bradley ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Typhoid Fever ◽  
Severe Disease ◽  
Genetically Engineered ◽  
Human Monoclonal Antibodies ◽  
Genetically Engineered Mice ◽  
Typhoid Toxin

ABSTRACT Typhoid toxin is a virulence factor of Salmonella enterica serovar Typhi, the causative agent of typhoid fever, and is thought to be responsible for the symptoms of severe disease. This toxin has a unique A2B5 architecture with two active subunits, the ADP ribosyl transferase PltA and the DNase CdtB, linked to a pentameric B subunit, which is alternatively made of PltB or PltC. Here, we describe the generation and characterization of typhoid toxin-neutralizing human monoclonal antibodies by immunizing genetically engineered mice that have a full set of human immunoglobulin variable region genes. We identified several monoclonal antibodies with strong in vitro and in vivo toxin-neutralizing activity and different mechanisms of toxin neutralization. These antibodies could serve as the basis for the development of novel therapeutic strategies against typhoid fever.

scholarly journals Monoclonal antibodies targeting the influenza virus N6 neuraminidase

2021 ◽  
Author(s):  
Shirin Strohmeier ◽  
Fatima Amanat ◽  
Juan Manuel Carreño ◽  
Florian Krammer
Keyword(s):  
Monoclonal Antibodies ◽  
Influenza A ◽  
Neutralization Assay ◽  
Escape Mutant ◽  
Influenza A Viruses ◽  
Highly Pathogenic ◽  
Lateral Ridge

AbstractInfluenza A viruses are a diverse species that include 16 hemagglutinin (HA) subtypes and 9 neuraminidase (NA) subtypes. While the antigenicity of many HA subtypes is reasonably well studied, less is known about NA antigenicity, especially when it comes to non-human subtypes that only circulate in animal reservoirs. The N6 NA subtypes are mostly found in viruses infecting birds. However, they have also been identified in viruses that infect mammals, such as swine and seals. More recently, highly pathogenic H5N6 subtype viruses have caused rare infections and mortality in humans. Here, we generated murine mAbs to the N6 NA, characterized their breadth and antiviral properties in vitro and in vivo and mapped their epitopes by generating escape mutant viruses. We found that the antibodies had broad reactivity across the American and Eurasian N6 lineages, but relatively little binding and inhibition of the H5N6 NA. Several of the antibodies exhibited strong NA inhibition activity and some also showed activity in the antibody dependent cellular cytotoxicity reporter assay and neutralization assay. In addition, we generated escape mutant viruses for six monoclonal antibodies and found mutations on the lateral ridge of the NA. Lastly, we observed variable protection in H4N6 and H5N6 mouse challenge models when the antibodies were given prophylactically.ImportanceThe N6 NA has recently gained prominence due to the emergence of highly pathogenic H5N6 viruses. Currently, there is limited characterization of the antigenicity of avian N6 neuraminidase. Our data is an important first step towards a better understanding of the N6 NA antigenicity.

999 In Vitro and In Vivo Characterization of Neutralizing Monoclonal Antibodies Against Clostridium difficile Toxins a and B

2013 ◽  
Vol 144 (5) ◽  
pp. S-185
Author(s):  
Dominiek Staelens ◽  
Marlies Van de Wouwer ◽  
Els Brouwers ◽  
Silvia Caluwaerts ◽  
Borden Lacy ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Clostridium Difficile ◽  
In Vivo Characterization

Immunolocalization and characterization of the low molecular weight antigen (4–5 kDa) ofToxoplasma gondiithat elicits an early IgM response upon primary infection

Parasitology ◽  
1994 ◽  
Vol 108 (2) ◽  
pp. 139-145 ◽  
Author(s):  
S. Tomavo ◽  
G. Couvreur ◽  
M. A. Leriche ◽  
A. Sadak ◽  
A. Achbarou ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Primary Infection ◽  
Low Molecular Weight ◽  
Immuno Electron Microscopy ◽  
Surface Localization

SUMMARYA striking feature of toxoplasmic seroconversion is the prominent and early IgM response to a low molecular weight antigen of 4–5 kDa. Two different monoclonal antibodies directed against the 4–5 kDa antigen have been generated and used to characterize this molecule. Using these monoclonal antibodies, we could demonstrate the surface localization of the lowMrantigen by immunofluorescence and immuno-electron microscopy assays. By immunoblotting, we observed that one of the monoclonal antibodies was unable to recognize the 4–5 kDa antigen in tachyzoites propagated in cell culture, indicating an epitope variability betweenToxoplasma gondiitachyzoites grownin vivoandin vitro. We discuss the implications of this latter finding in the design of diagnostic reagents.

Characterization of Therapeutic Monoclonal Antibodies Reveals Differences Between In Vitro and In Vivo Time-Course Studies

2012 ◽  
Vol 30 (1) ◽  
pp. 167-178 ◽  
Author(s):  
Sheng Yin ◽  
Cinthia V. Pastuskovas ◽  
Leslie A. Khawli ◽  
John T. Stults
Keyword(s):  
Monoclonal Antibodies ◽  
Time Course ◽  
Therapeutic Monoclonal Antibodies

scholarly journals Obtaining and Characterization of Hybridomas Producing Monoclonal Antibodies to Shiga-Like Toxins of I and II Types

2021 ◽  
pp. 83-88
Author(s):  
G. V. Kuklina ◽  
S. S. Ipatov ◽  
D. V. Pechenkin ◽  
A. V. Eremkin ◽  
A. A. Kytmanov ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Lines ◽  
Hybrid Cell ◽  
Sandwich Elisa ◽  
Minimum Detectable Concentration ◽  
Myeloma Cells ◽  
Hybrid Cell Lines

Objective – obtaining and characterization of hybrid cell lines producing monoclonal antibodies against I and II types of shiga-like toxins.Materials and methods. Shiga-like toxins obtained in “48thCentral Research Institute” of Ministry of Defense of Russian Federation (Kirov), BALB/c mice, myeloma cells SP2/0-Ag14 were used in research. Immune splenocytes and SP2/0-Ag14 myeloma cells were fused according to G. Kohler and C. Milstein method in De St. Fazekas and D. Scheidegger modifcation using 50 % polyethylene glycol. Hybrid cell lines producing specifc monoclonal antibodies were cloned by limited dilutions. Hybridomas growth and producing properties were studied in vitro and in vivo. Specifc activity of immune sera, culture and ascitic fluids were studied by indirect ELISA. Monoclonal antibodies from ascitic fluids were precipitated by saturated ammonium sulfate, followed by ion exchange chromatographyResults and discussion. 8 hybridomas producing monoclonal antibodies against I and II types shiga-like toxins were obtained. Hybridomas are characterized by stable proliferation and antibody-producing activity during 10 passages in vitro and 3 passages in vivo (observation period). Obtained monoclonal antibodies can be used for ELISA detection of I and II types shiga-like toxins. Minimum detectable concentration of shiga-like toxins in sandwich ELISA is 1 ng/ml. The possibility of detecting shiga-like toxins without typical differentiation was shown when using in the enzyme immunoassay a polyreceptor mixture of monoclonal antibodies for sensitizing the plate and a polyspecifc mixture of immunoperoxidase conjugates.

scholarly journals Isolation and characterization of human monoclonal antibodies to pneumococcal capsular polysaccharide 3

2021 ◽  
Author(s):  
Rachelle Babb ◽  
Christopher R Doyle ◽  
Liise-anne Pirofski
Keyword(s):  
Monoclonal Antibodies ◽  
Capsular Polysaccharide ◽  
Clinical Strain ◽  
Isolation And Characterization ◽  
Variable Heavy ◽  
Protective Antibodies ◽  
Intraperitoneal Infection

The current pneumococcal capsular polysaccharide (PPS) conjugate vaccine (PCV13) is less effective against Streptococcus pneumoniae serotype 3 (ST3), which remains a major cause of pneumococcal disease and mortality. Therefore, dissecting structure-function relationships of human PPS3 antibodies may reveal characteristics of protective antibodies. Using flow cytometry, we isolated PPS3-binding memory B cells from pneumococcal vaccine recipients and generated seven human PPS3-specific monoclonal antibodies (humAbs). Five humAbs displayed ST3 opsonophagocytic activity, four induced ST3 agglutination in vitro, and four mediated both activities. For two humAbs, C10 and C27, that used the same variable heavy (VH) and light (VL) chain domains (VH3-9*01/VL2-14*03), C10 had fewer VL somatic mutations, higher PPS3 affinity, more ST3 opsonophagocytic and agglutinating activity, whilst both humAbs altered ST3 gene expression in vitro. After VL swaps, C10VH/C27VL exhibited reduced ST3 binding and agglutination, but C27VH/C10VL binding was unchanged. In C57Bl/6 mice, C10 and C27 reduced nasopharyngeal colonization with ST3 A66 and a clinical strain, B2, and prolonged survival following lethal A66 intraperitoneal infection, but only C10 protected against lethal intranasal infection with the clinical strain. Our findings, associate efficacy of PPS3-specific humAbs with ST3 agglutination and opsonophagocytic activity and reveal an unexpected role for the VL in functional activity in vitro and in vivo. These findings also provide insights that may inform antibody-based therapy and identification of surrogates of vaccine efficacy against ST3.

A Molecular Approach to Immunoscintigraphy: A Study of the T-Antigen Conformation on the Surface of Tumors

1987 ◽  
Vol 26 (01) ◽  
pp. 1-6 ◽  
Author(s):  
S. Selvaraj ◽  
M. R. Suresh ◽  
G. McLean ◽  
D. Willans ◽  
C. Turner ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Tumor Cell ◽  
T Antigen ◽  
Human Carcinomas ◽  
Animal Tumor ◽  
Synthetic Antigens

The role of glycoconjugates in tumor cell differentiation has been well documented. We have examined the expression of the two anomers of the Thomsen-Friedenreich antigen on the surface of human, canine and murine tumor cell membranes both in vitro and in vivo. This has been accomplished through the synthesis of the disaccharide terminal residues in both a and ß configuration. Both entities were used to generate murine monoclonal antibodies which recognized the carbohydrate determinants. The determination of fine specificities of these antibodies was effected by means of cellular uptake, immunohistopathology and immunoscintigraphy. Examination of pathological specimens of human and canine tumor tissue indicated that the expressed antigen was in the β configuration. More than 89% of all human carcinomas tested expressed the antigen in the above anomeric form. The combination of synthetic antigens and monoclonal antibodies raised specifically against them provide us with invaluable tools for the study of tumor marker expression in humans and their respective animal tumor models.

Sign in / Sign up

Export Citation Format

Share Document