Several methods were applied to determine the viability of hybridoma cells in suspension. These methods include dye inclusion and exclusion assays such as the classical trypan blue exclusion assay, the propidium iodide (PI) exclusion assay and the fluorescein diacetate (FDA) inclusion assay. Furthermore, the relation was studied between release of lactate dehydrogenase (LDH) by hybridoma cells and their viability. Also the ATP content of the cells and cellular heterogeneity as measured with a flow cytometer were determined in relation to cellular viability. The dye inclusion and exclusion assays using trypan blue, FDA, PI were shown to be useful methods to determine cellular viability. With the FDA and PI methods it was possible to obtain additional information about cells which are in a transition state between viable and non-viable. The viability according to the scatter properties of the cells appears to reflect the overall condition of the cells, although interpretation of the results is difficult. Measurement of LDH release in the culture fluid or the cytoplasmic ATP content could not be used as parameters for cell viability.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/BF02540030 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Antigenic determinants underlying IgE-mediated anaphylaxis to peanut.
J Allergy Clin Immunol
January 2025
Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Vanderbilt University, Nashville, TN; Department of Pharmacology, Vanderbilt University Medical Center, Vanderbilt University, Nashville, TN.
Background: Studies of human IgE and its targeted epitopes on allergens have been very limited. We have an established method to immortalize IgE encoding B cells from allergic individuals.
Objective: To develop an unbiased and comprehensive panel of peanut-specific human IgE mAbs to characterize key immunodominant antigenic regions and epitopes on peanut allergens to map the molecular interactions responsible for inducing anaphylaxis.
Antigen Specificity: A Fluctuating Aspect in the Development of Clinical Antibodies?
APMIS
January 2025
Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Mohali, Punjab, India.
Development of antibodies for clinical use is a complex process involving numerous aspects, with antigen specificity being the most important. Initially, polyclonal antibodies, that can recognize multiple specific and nonspecific antigens (polyreactive), were developed and were very effective in the treatments. Later on, the polyspecificity/polyreactivity of these polyclonal antibodies (binding to multiple antigens) raised concerns about therapeutic efficacy because of their nonspecific interactions and challenges, such as development of immune complexes, batch-to-batch variability.
View Article and Find Full Text PDF[Preparation and identification of monoclonal antibodies against human LAG3 by immunizing mice with recombinant eukaryotic cell antigens].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
December 2024
Department of Medical Experimental Center, Northern Jiangsu People's Hospital, Yangzhou 225001, China. *Corresponding author, E-mail: yyue_king
Objective To prepare mouse anti-human lymphocyte activation gene 3 (LAG3) monoclonal antibody (mAb) and perform immunological identification of the antibody. Methods BALB/c mice were immunized with LAG3-mLumin-3T3 cells, which stably express the extracellular and transmembrane regions of human LAG3 in mouse 3T3 cells. The secretion of anti-human LAG3 antibodies in mouse serum was assessed using flow cytometry and immunofluorescence.
View Article and Find Full Text PDFA method for screening functional anti-Treg antibodies using a Treg-like cell line.
J Leukoc Biol
December 2024
School of Life Sciences, Arizona State University, Phoenix, AZ.
Regulatory T cells can suppress activated T cell proliferation by direct cell-contact, although the exact mechanism is poorly understood. Identification of a Treg-specific cell surface molecule that mediates suppression would offer a unique target for cancer immunotherapy to inhibit Treg immunosuppressive function or deplete Tregs in the tumor microenvironment. In this study, we explored a method of whole cell immunization using a Treg-like cell line (MoT cells) to generate and screen monoclonal antibodies that bound cell surface proteins in their native conformations and functionally reversed Treg-mediated suppression.
View Article and Find Full Text PDFThe Development of Monoclonal Antibody Against Thyroid-Stimulating Hormone for Congenital Hypothyroidism Screening in Indonesia.
Monoclon Antib Immunodiagn Immunother
December 2024
Endocrinology Division, Department of Pediatrics of Saiful Anwar General Hospital, Universitas Brawijaya, Malang, Indonesia.
Congenital hypothyroidism (CH) is a major health issue that can lead to intellectual disability if not detected and treated earlier. The preliminary screening program for neonatal CH in Indonesia gave a provisional incidence of 1:2513. Newborn screening using a dried blood spot sample is the standard method for CH detection, but it has limitations.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!