US 11,946,074 B2
Methods and compositions for inducible extracellular membrane capture of monoclonal immunoglobulins secreted by hybridomas
Scott K. Dessain, Wynnewood, PA (US); and Rama Devudu Puligedda, Secane, PA (US)
Assigned to Lankenau Institute for Medical Research, Wynnewood, PA (US)
Appl. No. 16/495,552
Filed by Lankenau Institute for Medical Research, Wynnewood, PA (US)
PCT Filed Mar. 23, 2018, PCT No. PCT/US2018/024073
§ 371(c)(1), (2) Date Sep. 19, 2019,
PCT Pub. No. WO2018/175917, PCT Pub. Date Sep. 27, 2018.
Claims priority of provisional application 62/526,608, filed on Jun. 29, 2017.
Claims priority of provisional application 62/476,599, filed on Mar. 24, 2017.
Prior Publication US 2020/0010810 A1, Jan. 9, 2020
Int. Cl. C12N 5/16 (2006.01); C07K 16/10 (2006.01); C12N 15/10 (2006.01); G01N 33/569 (2006.01)
CPC C12N 5/163 (2013.01) [C07K 16/10 (2013.01); C12N 15/1034 (2013.01); G01N 33/56966 (2013.01); C07K 2317/14 (2013.01); C07K 2317/21 (2013.01)] 7 Claims
 
1. A method of identifying a hybridoma cell that secretes a monoclonal antibody among a heterogeneous population of hybridoma cells, the method comprising:
(a) providing the hybridoma cells having been prepared by fusing B lineage cells with immortalized cells having expressed on their outer plasma membrane surfaces an Anchor, wherein said Anchor comprises i) an extracellular region comprising an antibody or fragment thereof capable of binding a selected Linker to form a first complex, and ii) a transmembrane amino acid sequence that immobilizes the Anchor on the outer plasma membrane surfaces, wherein said Anchor cannot bind said monoclonal antibody;
(b) contacting the hybridoma cells of (a) with an effective amount of said Linker, wherein said Linker is a soluble antibody or antibody fragment comprising a variable domain that binds and forms a second complex with a conserved domain of the monoclonal antibody, wherein the contacting permits the monoclonal antibody to be linked to the surface of the hybridoma cell that secretes it upon formation of said first and second complexes; and
(c) identifying the existence, antigen specificity, antigen binding affinity, titer, amount, or biological activity of the monoclonal antibody secreted by the hybridoma cells and bound thereto by said first and second immune complexes formed by Anchor-Linker-monoclonal antibody.