We describe a novel method for the production of monoclonal antibodies using a secretion capture report web (SCRW). Following HAT selection in bulk culture, individual hybridomas are encapsulated in biotinylated agarose drops. Antibody secreted by the hybridoma is captured within the agarose drop using an avidin bridge and biotinylated anti-mouse immunoglobulin. The secreted antibody is detected by a fluorescent reporter which can be either a second anti-mouse antibody or an antigen. The binding of the reporter can be quantitated and the desired hybridoma directly cloned by flow cytometry. Multiparameter (i.e., two-color) reporter analysis can also be used to selectively enrich and clone rare hybridomas secreting antibodies directed to unique epitopes. The method allows the characterization of thousands of clones per second and the isolation of hundreds of clones per day.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1038/nbt0895-787 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
SIV Env RhmAbs + N-803 at ART initiation prolongs viral decay without disrupting reservoir establishment in SIV-infected infant macaques.
PLoS Pathog
January 2025
Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, United States of America.
The latent viral reservoir remains the major barrier to HIV cure, placing the burden of strict adherence to antiretroviral therapy (ART) on people living with HIV to prevent recrudescence of viremia. For infants with perinatally acquired HIV, adherence is anticipated to be a lifelong need. In this study, we tested the hypothesis that administration of ART and viral Envelope-specific rhesus-derived IgG1 monoclonal antibodies (RhmAbs) with or without the IL-15 superagonist N-803 early in infection would limit viral reservoir establishment in SIV-infected infant rhesus macaques.
View Article and Find Full Text PDFExploring the Potential of Nanocarriers for Cancer Immunotherapy: Insights into Mechanism, Nanocarriers, and Regulatory Perspectives.
ACS Appl Bio Mater
January 2025
Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research, Hyderabad 500037, India.
Immunotherapy is a cutting-edge approach that leverages sophisticated technology to target tumor-specific antibodies and modulate the immune system to eradicate cancer and enhance patients' quality of life. Bioinformatics and genetic science advancements have made it possible to diagnose and treat cancer patients using immunotherapy technology. However, current immunotherapies against cancer have limited clinical benefits due to cancer-associated antigens, which often fail to interact with immune cells and exhibit insufficient therapeutic targeting with unintended side effects.
View Article and Find Full Text PDFRisk of infections in bispecific antibody therapy for multiple myeloma: a comprehensive review of literature.
Hematology
December 2025
Clinical Pharmacy Department, King Fahad Medical City, Riyadh, RH, Saudi Arabia.
Multiple Myeloma (MM) is a malignancy characterized by abnormal production of monoclonal immunoglobulins in plasma cells. Bispecific antibodies have emerged as a significant advancement in MM treatment, offering high effectiveness and specificity by targeting different antigens such as BCMA, CD38, and FcRH5. However, the risk of infection poses a major challenge in MM patients, which is thought to be influenced by various factors.
View Article and Find Full Text PDFEGFR-targeting RNase A-cetuximab antibody-drug conjugate induces ROS-mediated apoptosis to overcome drug resistance in KRAS mutant cancer cells.
Sci Rep
January 2025
Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz University of Medical Sciences, Tabriz, Iran.
Antibody-drug conjugates (ADCs) are an emerging strategy in cancer therapy, enhancing precision and efficacy by linking targeted antibodies to potent cytotoxic agents. This study introduces a novel ADC that combines ribonuclease A (RNase A) with cetuximab (Cet), an anti-EGFR monoclonal antibody, through a polyethylene glycol (PEG) linker (RN-PEG-Cet), aimed to induce apoptosis in KRAS mutant colorectal cancer (CRC) via a ROS-mediated pathway. RN-PEG-Cet was successfully synthesized and characterized for its physicochemical properties, retaining full enzymatic activity in RNA degradation and high binding affinity to EGFR.
View Article and Find Full Text PDFLeveraging prior knowledge for process parameter classification in mAb Protein A chromatography.
J Chromatogr A
December 2024
Eli Lilly and Company, Indianapolis, IN 46285, USA.
Protein A (ProA) affinity chromatography plays an essential role in purifying monoclonal antibodies (mAbs) and their analogues by reducing impurities like residual host cell proteins (HCPs), residual DNA, process additives, and potential viral contaminants. Decades of mAb process development and commercialization efforts have built extensive prior knowledge in the Protein A process. The prior knowledge facilities streamlined process development and minimized the need for extensive process characterization studies to inform manufacturing control strategies.
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!