Immunoglobulin Ihhibiting Reagent®: Evaluation of a New Method for Eliminating Spurious Elevations in CA125 Caused by HAMA

Cancer therapy utilising radiolabelled murine monoclonal antibodies frequently leads to the production of Human Anti-Mouse Antibodies (HAMA) in the recipient. HAMA interferes with “sandwich” immunoassays, such as those for tumour markers, rendering results unreliable. Published methods for eliminating HAMA from serum are not suitable for use in a laboratory which is processing a large number of assays using an automated system. We report on the use of Immunoglobulin Inhibiting Reagent (IIR) in CA125 assays from recipients of intraperitoneal radioimmunotherapy who had spuriously elevated results due to HAMA. IIR was found to be comparable to the admixture of mouse serum as a way of eliminating the effect of HAMA. IIR is ideally suited to use with an automated assay process.

Download Full-text

A novel modification of the avidin-biotin complex method for immunohistochemical studies of transgenic mice with murine monoclonal antibodies.

Journal of Histochemistry & Cytochemistry ◽

10.1177/40.9.1506669 ◽

1992 ◽

Vol 40 (9) ◽

pp. 1319-1328 ◽

Cited By ~ 23

Author(s):

K M Fung ◽

A Messing ◽

V M Lee ◽

J Q Trojanowski

Keyword(s):

Monoclonal Antibodies ◽

Transgenic Mice ◽

Detection System ◽

Immunohistochemical Method ◽

Mouse Tissue ◽

Mouse Serum ◽

Complex Method ◽

Mouse Tissues ◽

Secondary Antibodies ◽

Murine Monoclonal Antibodies

When mouse tissues are probed with murine monoclonal antibodies (MAb) by indirect immunohistochemistry, the secondary antibody detects tissue-bound MAb and irrelevant, endogenous mouse immunoglobulins. The latter are a source of confounding background, especially in diseased tissues. To circumvent this problem, we generated complexes of primary MAb and biotinylated secondary antibodies in vitro for use as antigen-specific probes. After blocking free binding sites in the complexed secondary antibodies with normal mouse serum, the complexes were applied to mouse tissue sections and tissue-bound complexes were visualized with an avidin-biotin detection system. Complexes formed with 12 different rat or mouse MAb were used to probe sections of normal mice, tumor-bearing transgenic mice, and mice with tumor xenografts. The staining patterns produced by these probes reflected the specificity of the MAb in the complexes, and the labeling of irrelevant, endogenous mouse immunoglobulins was reduced substantially. This novel, indirect immunohistochemical method can be exploited to study normal and diseased mouse tissues using a variety of murine MAb.

Download Full-text

Monoclonal Antibodies for Cancer Therapy Approved by FDA

MOJ Immunology ◽

10.15406/moji.2016.04.00120 ◽

2016 ◽

Vol 4 (2) ◽

Author(s):

Henry Hongrong Cai

Keyword(s):

Monoclonal Antibodies ◽

Cancer Therapy

Download Full-text

Monoclonal antibodies in contemporary cancer therapy

Monoclonal Antibodies in Oncology ◽

10.2217/ebo.13.127 ◽

2013 ◽

pp. 2-4

Author(s):

Fatih M Uckun

Keyword(s):

Monoclonal Antibodies ◽

Cancer Therapy

Download Full-text

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.

Download Full-text