Monoclonal Antibodies Monoclonal antibody for Glycans as Tools for Identifying Endogenous Glycan Ligands for Human Carbohydrate-Recognition Molecules

Fluorescent antibody technique by the monoclonal antibody method is very useful and helpful for the rapid quantification and in situ detection of the specific bacteria like nitrifiers in a mixed baxterial habitat such as a biofilm. In this study, twelve monoclonal antibodies against Nitrosomonas europaea (IFO14298) and sixteen against Nitrobacter winogradskyi (IFO14297) were raised from splenocytes of mice (BALB/c). It was found that these antibodies exhibited little cross reactivity against various kinds of heterotrophic bacteria. The direct cell count method using monoclonal antibodies could exactly detect and rapidly quantify N. europaea and N. winogradskyi. Moreover, the distribution of N. europaea and N. winogradskyi in a biofilm could be examined by in situ fluorescent antibody technique. It was shown that most of N. winogradskyi existed near the surface part and most of N. europaea existed at the inner part of the polyethylene glycol (PEG) gel pellet, which had entrapped activated sludge and used in a landfill leachate treatment reactor. It was suggested that this monoclonal antibody method was utilized for estimating and controlling the population of nitrifying bacteria as a quick and favorable tool.

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Therapeutic Monoclonal Antibodies in Clinical Practice against Cancer

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200703191653 ◽

2020 ◽

Vol 20 (16) ◽

pp. 1895-1907

Author(s):

Navgeet Kaur ◽

Anju Goyal ◽

Rakesh K. Sindhu

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Clinical Practice ◽

Therapeutic Agent ◽

Hematologic Malignancies ◽

Immune Checkpoints ◽

Malignant Cells ◽

Main Target ◽

Therapeutic Monoclonal Antibodies ◽

Cancerous Cells

The importance of monoclonal antibodies in oncology has increased drastically following the discovery of Milstein and Kohler. Since the first approval of the monoclonal antibody, i.e. Rituximab in 1997 by the FDA, there was a decline in further applications but this number has significantly increased over the last three decades for various therapeutic applications due to the lesser side effects in comparison to the traditional chemotherapy methods. Presently, numerous monoclonal antibodies have been approved and many are in queue for approval as a strong therapeutic agent for treating hematologic malignancies and solid tumors. The main target checkpoints for the monoclonal antibodies against cancer cells include EGFR, VEGF, CD and tyrosine kinase which are overexpressed in malignant cells. Other immune checkpoints like CTLA-4, PD-1 and PD-1 receptors targeted by the recently developed antibodies increase the capability of the immune system in destroying the cancerous cells. Here, in this review, the mechanism of action, uses and target points of the approved mAbs against cancer have been summarized.

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Evaluation of the Efficiency of Protein A Affinity Chromatography to Purify a Monoclonal Antibody for Cancer Treatment and its Purity Analysis

Current Chromatography ◽

10.2174/2213240607999201029204934 ◽

2020 ◽

Vol 7 (2) ◽

pp. 121-133

Author(s):

Ayesha Akhtar ◽

Shivakumar Arumugam ◽

Shoaib Alam

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Affinity Chromatography ◽

Protein A ◽

Exchange Chromatography ◽

Size Exclusion ◽

High Yield ◽

Staphylococcal Protein A ◽

Purification Step ◽

Purity Analysis

Background:: Protein A affinity chromatography is often employed as the most crucial purification step for monoclonal antibodies to achieve high yield with purity and throughput requirements. Introduction:: Protein A, also known as Staphylococcal protein A (SPA) is found in the cell wall of the bacteria staphylococcus aureus. It is one of the first discovered immunoglobulin binding molecules and has been extensively studied since the past few decades. The efficiency of Protein A affinity chromatography to purify a recombinant monoclonal antibody in a cell culture sample has been evaluated, which removes 99.0% of feed stream impurities. Materials and Method:: We have systematically evaluated the purification performance by using a battery of analytical methods SDS-PAGE (non-reduced and reduced sample), Cation Exchange Chromatography (CEX), Size-exclusion chromatography (SEC), and Reversed phased-Reduced Chromatography for a CHO-derived monoclonal antibody. Results and Discussion:: The analytical test was conducted to determine the impurity parameter, Host Cell Contaminating Proteins (HCP). It was evaluated to be 0.015ng/ml after the purification step; while initially, it was found to be 24.431ng/ml. Conclusion:: The tests showed a distinct decrease in the level of different impurities after the chromatography step. It can be concluded that Protein A chromatography is an efficient step in the purification of monoclonal antibodies.

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Development of a New Highly Selective Monoclonal Antibody against Preferentially Expressed Antigen in Melanoma (PRAME) and Identification of the Target Epitope by Bio-Layer Interferometry

International Journal of Molecular Sciences ◽

10.3390/ijms22063166 ◽

2021 ◽

Vol 22 (6) ◽

pp. 3166

Author(s):

Jwala Priyadarsini Sivaccumar ◽

Antonio Leonardi ◽

Emanuela Iaccarino ◽

Giusy Corvino ◽

Luca Sanguigno ◽

...

Keyword(s):

Mass Spectrometry ◽

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Western Blotting ◽

Cancer Biomarkers ◽

Trypsin Digestion ◽

Protein Fragments ◽

Target Epitope ◽

Potential Activity ◽

Antibody Epitopes

Background: Monoclonal antibodies (mAbs) against cancer biomarkers are key reagents in diagnosis and therapy. One such relevant biomarker is a preferentially expressed antigen in melanoma (PRAME) that is selectively expressed in many tumors. Knowing mAb’s epitope is of utmost importance for understanding the potential activity and therapeutic prospective of the reagents. Methods: We generated a mAb against PRAME immunizing mice with PRAME fragment 161–415; the affinity of the antibody for the protein was evaluated by ELISA and SPR, and its ability to detect the protein in cells was probed by cytofluorimetry and Western blotting experiments. The antibody epitope was identified immobilizing the mAb on bio-layer interferometry (BLI) sensor chip, capturing protein fragments obtained following trypsin digestion and performing mass spectrometry analyses. Results: A mAb against PRAME with an affinity of 35 pM was obtained and characterized. Its epitope on PRAME was localized on residues 202–212, taking advantage of the low volumes and lack of fluidics underlying the BLI settings. Conclusions: The new anti-PRAME mAb recognizes the folded protein on the surface of cell membranes suggesting that the antibody’s epitope is well exposed. BLI sensor chips can be used to identify antibody epitopes.

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A novel method for the non-chromatographic purification of technetium-99m-labeled monoclonal antibodies: a study with B72.3 monoclonal antibody

Nuclear Medicine and Biology ◽

10.1016/0969-8051(94)90006-x ◽

1994 ◽

Vol 21 (2) ◽

pp. 171-178 ◽

Cited By ~ 1

Author(s):

Howard S. Rosenzweig ◽

Girish N. Ranadive ◽

Troy Seskey ◽

Michael W. Epperly ◽

William D. Bloomer

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Chromatographic Purification ◽

Technetium 99M ◽

Novel Method

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Possible Future Monoclonal Antibody (mAb)-Based Therapy against Arbovirus Infections

BioMed Research International ◽

10.1155/2013/838491 ◽

2013 ◽

Vol 2013 ◽

pp. 1-21 ◽

Cited By ~ 12

Author(s):

Giuseppe Sautto ◽

Nicasio Mancini ◽

Giacomo Gorini ◽

Massimo Clementi ◽

Roberto Burioni

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Side Effects ◽

Antiviral Activity ◽

The Other ◽

Viral Escape ◽

Passive Immunotherapy ◽

Medical Need

More than 150 arboviruses belonging to different families are known to infect humans, causing endemic infections as well as epidemic outbreaks. Effective vaccines to limit the occurrence of some of these infections have been licensed, while for the others several new immunogens are under development mostly for their improvements concerning safety and effectiveness profiles. On the other hand, specific and effective antiviral drugs are not yet available, posing an urgent medical need in particular for emergency cases. Neutralizing monoclonal antibodies (mAbs) have been demonstrated to be effective in the treatment of several infectious diseases as well as in preliminaryin vitroandin vivomodels of arbovirus-related infections. Given their specific antiviral activity as well-tolerated molecules with limited side effects, mAbs could represent a new therapeutic approach for the development of an effective treatment, as well as useful tools in the study of the host-virus interplay and in the development of more effective immunogens. However, before their use as candidate therapeutics, possible hurdles (e.g., Ab-dependent enhancement of infection, occurrence of viral escape variants) must be carefully evaluated. In this review are described the main arboviruses infecting humans and candidate mAbs to be possibly used in a future passive immunotherapy.

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Monoclonal antibodies against chicken type IV and V collagens: electron microscopic mapping of the epitopes after rotary shadowing.

The Journal of Cell Biology ◽

10.1083/jcb.98.5.1637 ◽

1984 ◽

Vol 98 (5) ◽

pp. 1637-1644 ◽

Cited By ~ 32

Author(s):

R Mayne ◽

H Wiedemann ◽

M H Irwin ◽

R D Sanderson ◽

J M Fitch ◽

...

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Cleavage Site ◽

Type Iv Collagen ◽

Collagen Molecule ◽

Electron Microscopic ◽

Type V Collagen ◽

Type Iv ◽

Type V ◽

Rotary Shadowing

The location of the epitopes for monoclonal antibodies against chicken type IV and type V collagens were directly determined in the electron microscope after rotary shadowing of antibody/collagen mixtures. Three monoclonal antibodies against type IV collagen were examined, each one of which was previously demonstrated to be specific for only one of the three pepsin-resistant fragments of the molecule. The three native fragments were designated (F1)2F2, F3, and 7S, and the antibodies that specifically recognize each fragment were called, respectively, IA8 , IIB12 , and ID2 . By electron microscopy, monoclonal antibody IA8 recognized an epitope located in the center of fragment (F1)2F2 and in tetramers of type IV collagen at a distance of 288 nm from the 7S domain, the region of overlap of four type IV molecules. Monoclonal antibody IIB12 , in contrast, recognized an epitope located only 73 nm from the 7S domain. This result therefore provides direct visual evidence that the F3 fragment is located closest to the 7S domain and the order of the fragments must be 7S-F3-(F1)2F2. The epitope for antibody ID2 was located in the overlap region of the 7S domain, and often several antibody molecules were observed to binding to a single 7S domain. The high frequency with which antibody molecules were observed to bind to fragments of type IV collagen suggests that there is a single population of type IV molecules of chain organization [alpha 1(IV)]2 alpha 2(IV), and that four identical molecules must form a tetramer that is joined in an antiparallel manner at the 7S domain. The monoclonal antibodies against type V collagen, called AB12 and DH2 , were both found to recognize epitopes close to one another, the epitopes being located 45-48 nm from one end of the type V collagen molecule. The significance of this result still remains uncertain, but suggests that this site is probably highly immunoreactive. It may also be related to the specific cleavage site of type V collagen by selected metalloproteinases and by alpha-thrombin. This cleavage site is also known to be located close to one end of the type V molecule.

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Unreliable Measurement of Basophil Maximum Leukotriene Release with the Bühlmann CAST 2000 Enzyme-Linked Immunosorbent Assay Kit

Clinical and Vaccine Immunology ◽

10.1128/cvi.13.3.420-422.2006 ◽

2006 ◽

Vol 13 (3) ◽

pp. 420-422 ◽

Cited By ~ 6

Author(s):

S. E. Burastero ◽

C. Paolucci ◽

D. Breda ◽

G. Monasterolo ◽

R. E. Rossi ◽

...

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Immunosorbent Assay ◽

False Positive ◽

Positive Control ◽

Enzyme Linked Immunosorbent Assay ◽

Positive Results

ABSTRACT The Bühlmann CAST 2000 enzyme-linked immunosorbent assay is a potentially useful assay for measuring sulfidoleukotrienes released in vitro by allergen-challenged basophils. However, we observed that the positive-control reagent yielded positive signals in cell-free systems. These false-positive results depended on using a mouse anti-FcεRI monoclonal antibody and were prevented by degranulation-inducing reagents other than mouse monoclonal antibodies.

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Changes in expression of monoclonal antibody epitopes on laminin-5r induced by cell contact

Journal of Cell Science ◽

10.1242/jcs.109.7.1965 ◽

1996 ◽

Vol 109 (7) ◽

pp. 1965-1973 ◽

Cited By ~ 1

Author(s):

G. Plopper ◽

J. Falk-Marzillier ◽

S. Glaser ◽

M. Fitchmun ◽

G. Giannelli ◽

...

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Epithelial Cell ◽

Epithelial Cell Line ◽

Cell Contact ◽

Membrane Component ◽

Human Epithelial Cell ◽

Matrix Assembly ◽

Basement Membrane Component ◽

Antibody Epitopes

Laminin-5r is a basement membrane component that promotes rapid adhesion and hemidesmosome formation in epithelial cells. We raised monoclonal antibodies and identified their corresponding epitopes on the constituent chains of laminin-5r by western blotting. Using a combination of immunoprecipitation and ELISA assays, we determined that these epitopes are differentially exposed on two forms of the laminin-5r heterotrimer: soluble (passively adsorbed onto plastic) and cell-associated. Antibody 5C5 epitope is exposed on the cell-associated form, but not the soluble/passively adsorbed form of laminin-5r. Epitopes reactive with antibodies CM6, FM3, and TR1 are also preferentially exposed on cell-associated laminin-5r, such that reactivity of these antibodies with the cell-associated form is fourfold higher than with the soluble/passively adsorbed form in ELISA assays. Incubation of passively adsorbed laminin-5r with the human epithelial cell line SCC12 induced exposure of 5C5 and CM6, FM3, or TR1 epitopes. These data suggest that cells actively modify laminin-5r, perhaps during matrix assembly, and that the 5C5 epitope may serve as a marker for assembled laminin-5r matrix.

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

Revista da Sociedade Brasileira de Medicina Tropical ◽

10.1590/s0037-86821996000500012 ◽

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.

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