Process for producing donor-batched cells expressing a recombinant receptor

A method of preparing a T cell composition from a donor pool, the method comprising:

• (a) selecting T cells surface negative for CD57 (CD57−) from a donor sample from an individual donor, thereby generating a CD57 depleted T cell population;
• (b) genetically engineering the CD57 depleted T cell population, thereby producing an engineered T cell composition, the genetic engineering comprising:
• (1) knocking out expression of (i) an endogenous major histocompatibility complex (MHC) or a component thereof;
• and/or (ii) an endogenous T cell receptor (TCR) or a component thereof, in cells of the CD57 depleted T cell population;
• and
• (2) introducing a heterologous polynucleotide encoding a recombinant receptor into the cells of the CD57 depleted T cell population, wherein the recombinant receptor is a chimeric antigen receptor (CAR);
• wherein the knocking out in (1) and the introducing in (2) are carried out concurrently or successively in either order;
• (c) repeating steps (a) and (b) for a plurality of different donors to produce a plurality of donor engineered T cell compositions, wherein each donor engineered T cell composition is generated from cells from the donor sample from the individual donor of the plurality of different donors;
• and
• (d) combining the plurality of donor engineered T cell compositions from the plurality of different individual donors to produce a donor pooled engineered T cell composition.

A method of preparing a T cell composition from a donor pool, the method comprising:

• (i) selecting for one of (a) cells surface positive for a T cell marker(s) and (b) cells surface negative for CD57 (CD57−) from a donor sample from a plurality of different donors, thereby generating an enriched population of cells;
• and
• (ii) selecting, from the enriched population of cells, the other of (a) cells surface positive for the T cell marker(s) and (b) CD57− cells, thereby generating a CD57 depleted T cell population, wherein:
• (1) the donor sample is a pooled sample comprising cells from the plurality of different donors, whereby the method produces a pooled CD57 depleted T cell population;
• or
• (2) the donor sample is a sample from an individual donor, and steps (i) and (ii) are repeated separately for each donor sample from the plurality of different donors, whereby the method produces a CD57 depleted T cell population for each individual donor.