Table 3.

Self-association properties of insulin analogs

Analog	t1/2 Hexamer dissociation* (min ± SD)	Calculated molecular mass by SEC† (kDa ± fit error)
Human insulin (ins)	7.7 ± 1.3	6.8 ± 3.1
Ins lispro‡	4.6 ± 0.3	4.2 ± 2.0
OrnB29 -ins§	8.6 ± 1.3	6.6 ± 3.0
TyrB24, OrnB29-ins	7.6 ± 0.3	4.7 ± 2.2

^*t_1/2 and SD were calculated from three replicates; two-sided P values for lispro, Orn^B29-ins, and Tyr^B24, Orn^B29-ins against t_1/2 of human insulin were 0.06, 0.44, and 0.91, respectively.

^†Proteins were made 0.6 mM in a buffer containing ZnCl₂ at a ratio of two zinc ions per insulin hexamer and applied to SEC column (Materials and Methods); masses were calculated based on protein standards (SI Appendix, Fig. S19B).

^‡Insulin lispro (KP-insulin) contains substitutions Pro^B28→Lys (K) and Lys^B29→Pro (P).

^§Substitution Lys^B29→Orn was introduced for synthetic convenience.