CD19+ B lymphocytes are the major source of human antibody-secreting hybridomas generated by electrofusion

Schmidt, Enno; Leinfelder, Ulrich; Geßner, P.; Zillikens, Detlef; Bröcker, Eva‐Bettina; Zimmermann, U.

doi:10.1016/s0022-1759(01)00431-8

Cited by 20 publications

(8 citation statements)

References 27 publications

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“…1, step 4). MNCs were stimulated for 3 to 5 days with phytohemagglutinin-L (PHA-L) (Roche, Indianapolis, IN, 2.5 μg/ml) at 37°C/5% CO 2 before fusion (Schmidt et al, 2001). MNCs were fused with murine plasmacytoma P3X63.Ag8.653 (ATCC, Manassas, VA) (Kearney et al, 1979) under hypo-osmolar condition (Zimmermann et al, 1990) using a Multiporator/Helix chamber (Eppendorf, Westbury, NY).…”

Section: Hybridoma Generation and Evaluation Of Antibodiesmentioning

confidence: 99%

Autologous antibodies that bind neuroblastoma cells

Sun

Sholler

Shukla

et al. 2015

Journal of Immunological Methods

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Antibody therapy of neuroblastoma is promising and our goal is to derive antibodies from patients with neuroblastoma for developing new therapeutic antibodies. The feasibility of using residual bone marrow obtained for clinical indications as a source of tumor cells and a source of antibodies was assessed. From marrow samples, neuroblastoma cells were recovered, grown in cell culture and also implanted into mice to create xenografts. Mononuclear cells from the marrow were used as a source to generate phage display antibody libraries and also hybridomas. Growth of neuroblastoma patient cells was possible both in vitro and as xenografts. Antibodies from the phage libraries and from the monoclonal hybridomas bound autologous neuroblastoma cells with some selectivity. It appears feasible to recover neuroblastoma cells from residual marrow specimens and to generate human antibodies that bind autologous neuroblastoma cells. Expansion of this approach is underway to collect more specimens, optimize methods to generate antibodies, and to evaluate the bioactivity of neuroblastoma-binding antibodies.

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Section: Hybridoma Generation and Evaluation Of Antibodiesmentioning

confidence: 99%

Autologous antibodies that bind neuroblastoma cells

Sun

Sholler

Shukla

et al. 2015

Journal of Immunological Methods

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“…Mononuclear cells were stimulated for 3 to 5 days with phytohemagglutinin-L (PHA-L, 2.5 µg/ml) at 37°C/5% CO 2 before fusion [4]. The hybridoma fusion partner to B cells was murine plasmacytoma P3X63.Ag8.653 [5].…”

Section: Body Of Workmentioning

confidence: 99%

Marrow-Derived Antibody Library for Treatment of Neuroblastoma

Sholler¹

2013

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Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES)Van Andel Research Institute Grand Rapids, MI 49503 PERFORMING ORGANIZATION REPORT NUMBER SPONSORING / MONITORING AGENCY NAME(S) AND ADDRESS(ES) 10. SPONSOR/MONITOR'S ACRONYM(S) U.S. Army Medical Research and Materiel Command Fort Detrick, Maryland 21702-5012 SPONSOR/MONITOR'S REPORT NUMBER(S) DISTRIBUTION / AVAILABILITY STATEMENTApproved for Public Release; Distribution Unlimited SUPPLEMENTARY NOTES ABSTRACTNeuroblastoma (NB) is the most common solid tumor in children, which accounts for 15% of all pediatric cancer deaths in the US. New therapeutic antibodies to treat NB are urgently needed to improve survival. The purpose of this project is to produce patient-specific therapeutic antibodies to treat neuroblastoma. The goal is to develop new methods and strategies to capture the auto-immune response reaction in neuroblastoma patients using phage display and B cell hybridoma technologies.

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“…The hybridoma is then able to continue to grow in culture, and a large amount of specific desired antibodies can be recovered after processing. Electrofusion has proved to be a successful approach in the production of vaccines Orentas et al (2001);Scott-Taylor et al (2000), antibodies Schmidt et al (2001), and reconstructed embryos in mammalian cloning Gaynor et al (2005).…”

Section: Electrofusion (Ef)mentioning

confidence: 99%

Therapeutic Applications of Electroporation

Talele¹

2012

Biotechnology - Molecular Studies and Novel Applications for Improved Quality of Human Life

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CD19+ B lymphocytes are the major source of human antibody-secreting hybridomas generated by electrofusion

Cited by 20 publications

References 27 publications

Autologous antibodies that bind neuroblastoma cells

Autologous antibodies that bind neuroblastoma cells

Marrow-Derived Antibody Library for Treatment of Neuroblastoma

Therapeutic Applications of Electroporation

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