TABLE 5

In vitro analytical data for hyperglycosylation variants of FS315-hFc

The recombinant FS315-hFc variants with newly designed one or two NXT/Ss are listed in the table. The binding of the variants to heparin, myostatin, or hFcRn was determined by SPR. The binding affinities were measured and reported by the KD. The charge heterogeneity of the variants was determined by cIEF and shown as the range of pI values. The statistical parameters and analysis, including association constant (ka) with S.E., dissociation constant (kd) with S.E., and the average KD values with S.D. (n = 4 independent batches) for an internal control used in all SPR assays, are reported in Supplemental Table 4.

FS315-hFc VariantHeparin Binding KDMyostatin Binding KDhFcRn Binding KDcIEF
nMpMnMpI
Wild type0.220.228.35.07–5.89
K82T1.415.013.85.29–5.93
P85T0.412.420.15.51–6.09
R78N/N80T0.913.020.15.47–6.09
R86N/V88T1.512.710.85.49–6.08
G74N/K76S0.311.672.85.06–5.88
G74N/K76T0.511.143.05.06–5.99
G74N/K76T/P85T0.311.629.64.86–5.87
K75N/C77T/K82T3.540.336.24.72–5.88
C66A/K75N/C77T0.324.427.54.81–6.47
C66S/K75N/C77T0.426.082.84.86–6.47