TABLE 3.

Sizes of single-chain constructs of BAFF and APRIL

Constructsa	Theoretical no. of N-linkedb	Theoretical approximate mass (+N-linked)c	Observed approximate mass, WB + DTT	Observed approximate mass, S200	Ratio, S200/WB
		kDa	kDa	kDa
FLAG-AAA	6	64.3	58	85	1.5
FLAG-BAA	5	62.5	60	90	1.5
FLAG-ABB	4	60.9	59	100	1.7
FLAG-BBB	3	59.3	58	95	1.6
Fc-AAA	7	92.7	98	270	2.8
Fc-BAA	6	91	98	330	3.4
Fc-ABB	5	89.4	93	500	5.4
Fc-BBB	4	88	100	500	5.0
BAA, ABA, AAB	3	56.1	51d	40	0.8
ABB, ABA, BBA	3	56.8	54e	45	0.8

^a Tagged constructs were produced in CHO cells, and untagged constructs were produced in HEK 293 cells.

^b Number of consensus N-glycosylation sites (NX(S/T)) is shown.

^c We assume that all consensus sites are glycosylated and counted 2.5 kDa per N-linked glycan.

^d 51- and 54-kDa bands were observed and are probably N-glycosylation species.

^e 51-, 54-, and 57-kDa bands were observed and are probably N-glycosylation species.