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UK Health Security Agency

55-6

55-6

Catalogue No.

97040308

Cell Line Name

55-6

Cell Line Description

The hybridoma cell line 55-6 produces antibodies reacting with the Human Immunodeficiency Virus I (HIV-1) envelope protein, gp120. Spleen cells from BALB/c mice that had been sequentially immunised with recombinant gp120(rgp120) from three different nonglycosylated isolates of HIV-1 (IIIB,SF2 and Z6) have been fused with Sp2/0-Ag14 myeloma cells. The antibody binds rgp120 from the III B isolate and reacts weakly with gp120 from the MN strain, both shown by ELISA. No cross blocking of any other antibodies in the developed panel was detected, thus indicating that 55-6 binds to a unique site on gp120IIIB. In peptide recognition assays it was shown to bind a synthetic peptide 105-117 (in the CI region) of gp120.

Characteristics

Myeloma

Sp2/0 Ag14

Immunogen

RGP 120

Immunological Donor

BALB/c mouse spleen

Antibody Isotype

IgG2a

Morphology

Spherical

Tissue of Origin

Hybrid

Culture Conditions

Cell Type

Hybridoma

Subculture Routine

Maintain cultures between 1-9 x 100,000 cells/ml; 5% CO₂;37°C. When recovering hybridoma cultures from frozen it is not unusual for growth to be slower than expected initially and there may be an observed decrease in viability. Establishment of an actively proliferating culture may take up to 2 weeks. On resuscitation a centrifugation step to remove the cryoprotectant is essential. Rapidly thaw the frozen ampoule in a water bath at 37°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add 5-10ml of prewarmed growth media+. Remove a sample for counting. Centrifuge at 100 x g for 2-3 minutes to pellet cells and seed at a relatively high density of 5-7 x 10µ cells/ml. Place culture flask flat and observe regularly until viable proliferating cells are seen. Often hybridoma cultures may benefit from being resuspended with media supplemented with 20% FBS in the early critical stage of culture establishment immediately post resuscitation. Once the culture is established in culture the FBS can be reduced to 10%.

Culture Medium

RPMI 1640 + 2mM glutamine + HT + 10% FBS

Growth Mode

Suspension

Additional Info

Depositor

Dr. S.L. Epstein, Laboratory of Immunology and Developmental Biology, Division of Cellular and Gene Therapies, CBER, FDA, USA

Country of Origin

United States

Applications

gp120 assays

Assays

ELISA, immunofluorescence cross-blocking assay, peptide ELISA

Hazard Group (ACDP)

2

References

References

Hybridoma 1995;14:235

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: 55-6 (ECACC 97040308)

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.