Generation of mid‐region murine monoclonal antibodies against multiple structural isoforms of amyloid‐beta by hybridoma technology

Many pathological conditions are accompanied with changes in the concentration of the total IgG or some of its fraction. For this reason there is great interest in the production of reagents specific for IgG. In this paper, the binding characteristics of two new murine monoclonal antibodies (MoAb), assigned MoAb 15 and MoAb 22, are reported. These MoAbs were produced by hybridoma technology. By performing ELISAs and Western blots analyzes, it was demonstrated that both MoAbs interact specifically with human IgG. Cross reactivity with other sera proteins was not observed. In order to precisely localize the epitopes recognized by MoAb 15 and MoAb 22, the Western blots interactions of these MoAbs with electrophoreticaly separated IgG-fragments, obtained by the action of proteolytic enzymes (papain, pepsin, trypsin), were analyzed. According to the results of these experiments, both MoAbs interacted with epitopes in the C 3 domain. The affinity constants, calculated from Scatchard plots of binding ofMoAb15 and MoAb22 to human IgG, wereKa15 = 1.71 106M-1 andKa22 = 2.15 109M-1. According to all these findings,MoAb 15 and MoAb 22 could be used in standard immunochemical techniques. However, the experiments showed that both MoAbs had bad immunoprecipitating properties. In solid phase techniques (ELISAs, Western blot, dot-blot, etc.), their application gave excellent results that highly recommended them for use in these types of analyzes.

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Preface hybridoma technology I: Murine monoclonal antibodies

Journal of Tissue Culture Methods ◽

10.1007/bf01665916 ◽

1985 ◽

Vol 9 (3) ◽

pp. 129-129 ◽

Cited By ~ 6

Author(s):

Elizabeth M. Earley ◽

Joan C. Rener

Keyword(s):

Monoclonal Antibodies ◽

Hybridoma Technology ◽

Murine Monoclonal Antibodies

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PERSPECTIVES OF APPLICATION OF ANTIBODIES AGAINST ENDOGLIN (CD105) FOR VISUALIZATION AND ANTI-ANGIOGENE THERAPY OF TUMORS

Problems in oncology ◽

10.37469/0507-3758-2018-64-4-504-507 ◽

2018 ◽

Vol 64 (4) ◽

pp. 504-507

Author(s):

Vladimir Klimovich ◽

Natalya Vartanyan ◽

Anastasiya Stolbovaya ◽

Lidiya Terekhina ◽

Olga Shashkova ◽

...

Keyword(s):

Endothelial Cells ◽

Monoclonal Antibodies ◽

Human Plasma ◽

Vascular Endothelium ◽

Immune Complexes ◽

Targeted Delivery ◽

Biological Fluids ◽

Therapeutic Drugs ◽

Cultured Endothelial Cells ◽

Hybridoma Technology

During last years monoclonal antibodies (MAB) directed against vascular endothelium markers demonstrated their efficiency for visualization and targeted delivery of therapeutic drugs to tumors. Endoglin (CD105) which serves as a key element that determines endothelial cells quiescence or activation is one of such markers. Endoglin is highly expressed on the vascular endothelium of growing tumors. A first panel of MAB against endoglin in our country was produced at the hybridoma technology laboratory of RRC RST named after A.M. Granov. On the basis of these MAB ELISA was created allowing detection of endoglin in human plasma and other biological fluids. Several MAB had been shown to bind endoglin on the membrane of the cultured endothelial cells and to persist there for several hours. During the first 30 min after binding some of the immune complexes “endoglin-MAB” were internalized into the cytoplasm and were found included in the endosomes. In future these MAB can be used to create the reagents for the addressed delivery of isotope tags both on the membrane and into the cytoplasm of endothelial cells.

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IMMUNOREACTIVITY OF NEURONAL LIPOFUSCIN WITH MONOCLONAL ANTIBODIES TO THE AMYLOID BETA PROTEIN

Alzheimer Disease & Associated Disorders ◽

10.1097/00002093-198802030-00111 ◽

1988 ◽

Vol 2 (3) ◽

pp. 262 ◽

Cited By ~ 2

Author(s):

C. BANCHER ◽

I. GRU NDKE-IQBAL ◽

K. IQBAL ◽

K. S. KIM ◽

H. M. WISNIEWSKI

Keyword(s):

Monoclonal Antibodies ◽

Amyloid Beta ◽

Amyloid Beta Protein

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Screening and development of monoclonal antibodies for identification of ferret T follicular helper cells

Scientific Reports ◽

10.1038/s41598-021-81389-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Wenbo Jiang ◽

Julius Wong ◽

Hyon-Xhi Tan ◽

Hannah G. Kelly ◽

Paul G. Whitney ◽

...

Keyword(s):

Animal Model ◽

Monoclonal Antibodies ◽

Lymph Node ◽

Cross Reactivity ◽

Tfh Cells ◽

Follicular Helper ◽

Lymph Node Cells ◽

Human Viruses ◽

Humoral Responses ◽

Murine Monoclonal Antibodies

AbstractThe ferret is a key animal model for investigating the pathogenicity and transmissibility of important human viruses, and for the pre‐clinical assessment of vaccines. However, relatively little is known about the ferret immune system, due in part to a paucity of ferret‐reactive reagents. In particular, T follicular helper (Tfh) cells are critical in the generation of effective humoral responses in humans, mice and other animal models but to date it has not been possible to identify Tfh in ferrets. Here, we describe the screening and development of ferret-reactive BCL6, CXCR5 and PD-1 monoclonal antibodies. We found two commercial anti-BCL6 antibodies (clone K112-91 and clone IG191E/A8) had cross-reactivity with lymph node cells from influenza-infected ferrets. We next developed two murine monoclonal antibodies against ferret CXCR5 (clone feX5-C05) and PD-1 (clone fePD-CL1) using a single B cell PCR-based method. We were able to clearly identify Tfh cells in lymph nodes from influenza infected ferrets using these antibodies. The development of ferret Tfh marker antibodies and the identification of ferret Tfh cells will assist the evaluation of vaccine-induced Tfh responses in the ferret model and the design of novel vaccines against the infection of influenza and other viruses, including SARS-CoV2.

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