Table 1.

Sequence summary of the Shuffle and ShuffleStEP libraries

			Mutational frequency (%)
Library	Round	[ARG2] (nM)	H1	H2	H3	L1	L2	L3	Recombination frequency (%)	CDR diversity (%)
Shuffle	0	N/A	32	43	32	30	44	33	90	100
ShuffleStEP		N/A	34	39	30	28	38	33	81	100*
Shuffle	2	5.0	63	43	3	13	43	35	70	100
ShuffleStEP		5.0	63	38	0	13	43	35	70	100
Shuffle	3	1.0	65	43	13	28	23	40	85	93
ShuffleStEP		1.0	64	51	8	23	33	28	74	100
Shuffle	3	0.5	50	65	5	13	33	63	85	90
ShuffleStEP		0.5	60	45	7	21	29	43	69	93

The table shows the abundance of clones with mutations in each CDR, as well as the recombination frequency and CDR amino acid diversity for each round. Mutational frequency is calculated by dividing the number of clones with amino acid mutations in the designated CDR by the number of clones sampled × 100. Recombination frequency was defined as the percentage of clones in the population with recombined mutations in different CDRs (number of clones with amino acid mutations in more than one CDR/number of clones sampled × 100). CDR diversity describes the percentage of clones in the population with a unique CDR sequence combination (number of clones with a unique CDR amino acid sequence combination/number of clones sampled × 100). The concentration of ARG2 used in each selection round is also indicated.

^*Excluding 2.5% parent.