Table 1.

Activity profiles of anti-human serum albumin (HSA) antibodies. Fluorescent microvolume assay technology (FMAT) screening of secreted anti-HSA antibodies in B cell supernatants for binding to 100ng/ml HSA in the presence or absence of 25 µM albumin binding compounds (warfarin, ibuprofen, myristic acid, and copper chloride) and for binding to 100ng/ml rat serum albumin (RSA). FL = fluorescence intensity. Equilibrium binding constants (K_D) of anti-HSA rabbit Fab fragments for human and mouse serum albumin (MSA), and of equivalent humanized IgG antibodies for HSA, MSA and RSA determined by surface plasmon resonance (SPR).

	FMAT			SPR
	B cell sup			Rabbit     Fab		Humanized
mAb CA#	HSA (FL)	HSA + compounds (FL)	RSA (FL)	HSA KD x10−9 (M)	MSA KD x10−9 (M)	HSA KD x10−9 (M)	MSA KD x10−9 (M)	IgGRSA KD x10−9 (M)	Yield (mg/ml)
645	272	220	114	0.31	2.6	0.82	2.9	7.9	161
646	2310	964	484	0.14	1.6	0.57	1.7	4.5	35
647	1213	520	69	0.60	36.0	1.30	26	10	23
648	1048	465	72	0.33	12.0	0.13	23	54	312
649	1338	534	142	0.54	13.0	0.32	17	44	188