Table 1 (abstract P-135). Enzymatic reactions were conducted in 25 mM Tris + 50 mM NaCl buffer at the specified pH for 48-72 h. The % yields of the targeted glycan structures are shown. FG0 = fucosylated agalactosylated, FG2 = fucosylated digalactosylated, FM3 = fucosylated trimannosyl core, FA1 = fucosylated mannosyl core plus one GlcNAc, FS1 = fucosylated monosialylated biantennary, FS2 = fucosylated disialylated biantennary

Agalactosylation

1

3 units/μL β(1,4) galactosidase, 3 units/μL neuraminidase at pH 6.6

EG2

FG0

85

Cetux

FG0

94

IL8

FG0

83

Polyclonal

FG0

62

Galactosylation

1

2 milli-units/μL β(1,4) galactosyltransferase, 3 mM UDP-galactose, 10 mM MnCl₂, 3 units/μL neuraminidase at pH7.5

EG2

FG2

96

Cetux

FG2

95

IL8

FG2

95

serum

FG2

96

Trimannose

(FM3 + FA1)

2

Stage 1: 3 units/μL β(1,4) galactosidase, 3 units/μL neuraminidase at pH6.6

Stage 2: 20 units of β-N-acetylglucosaminidase at pH6.6

EG2

FM3

80

FA1

11

Cetux

FM3

78

FA1

15

IL8

FM3

76

FA1

18

Sialylation

(FS1 + FS2)

2

Stage 1: 2 milli-units/μL β(1,4) galactosyltransferase, 3 mM UDP-galactose, 10 mM MnCl₂ at pH7.5

Stage 2: 100 μg/mL α(2,6) sialyltransferase, 3 mM CMP-Neu5Ac at pH7.5

EG2

FS2

76

FS1

16

IL8

FS2

37

FS1

38

Polyclonal

FS2

43

FS1

46