Table 1. Molecular masses and yields of T cell-engaging triplebodies and bispecific scFvs.

Proteins were expressed and purified as described in the Methods section. Buffer conditions were chosen to minimize protein aggregation. Proteins were concentrated to 150 - 300 ng/μL in the storage buffer by centrifugation, using centrifugal filters with a molecular weight cut-off (MWCO) of 50 kDa for the triplebodies and 10 kDa for bispecific scFvs. Theoretical molecular masses were computed from the known amino acid sequence composition of the molecules.

Protein	Computed Mass [kDa]	Yield [mg/L] (293F cells)	Storage Buffer
anti[CD19-CD3]	60.7	0.9 – 4.8	20mM L-Histidin 300mM NaCl 100mM D-Trehalose 5mM EDTA 10% Glycerol pH 6.5
anti[CD19-CD3-CD19]	89.9	0.9 – 5.1
anti[Her2-CD3]	59.5	1.8 – 9.2
anti[Her2-CD3-Her2]	87.2	6.0 – 8.5