Process for the modification of a glycoprotein using a glycosyltransferase that is or is derived from A β(1,4)-N-acetylgalactosaminyltransferase

1. A process for the modification of a glycoprotein, the process comprising:

contacting a glycoprotein comprising a glycan comprising a terminal GlcNAc moiety, with a sugar-derivative nucleotide Su(A)-Nuc in the presence of a glycosyltransferase,

wherein:

(i) the glycosyltransferase is, or is derived from, Ascaris summ β-(1,4)-N-acetylgalactosaminytransferase or Trichoplusia ni β-(1,4)-N-acetylgalactosaminyltransferase;

(ii) the glycan, comprising a terminal GlcNAc-moiety, is according to formula (1) and optionally (2)

wherein:

b is 0 or 1;

d is 0 or 1;

e is 0 or 1; and

G is a monosaccharide, or a linear or branched oligosaccharide comprising 2 to 20 sugar moieties; and

(iii) the sugar-derivative nucleotide Su(A)-Nuc is according to formula (3):

wherein:

a is 0 or 1;

f is 0 or 1;

g is 0 or 1;

Nuc is a nucleotide;

U is [C(R 1 ) 2 ]n or [C(R 1 ) 2 ] p —O—[C(R 1 ) 2 C(R 1 ) 2 O] o —[C(R 1 ) 2 ] q , wherein n is an integer in the range of 1 to 24; o is an integer in the range of 0 to 12; p and q are independently 0, 1 or 2; and R 1 is independently selected from the group consisting of H, F, Cl, Br, I, OH and an unsubstituted or substituted C 1 -C 24 alkyl group;

T is a C 3 -C 12 (hetero)arylene group, wherein the (hetero)arylene group is optionally substituted;

A is selected from the group consisting of:

(a) —N 3

(b) —C(O)R 3

wherein R 3 is an unsubstituted or substituted C 1 -C 24 alkyl group;

(c) (hetero)cycloalkynyl group or a (CH 2 ) i C≡C—R 4 moiety

wherein i is 0-10 and R 4 is hydrogen or an unsubstituted or substituted C 1 -C 24 alkyl group;

(d) —SH

(e) —SC(O)R 8

wherein R 8 is an, unsubstituted or substituted, C 1 -C 24 alkyl group or phenyl group;

(f) —SC(V)OR 8

wherein V is O or S, and R 8 is an, unsubstituted or substituted, C 1 -C 24 alkyl group or phenyl group;

(g) —X

wherein X is selected from the group consisting of F, Cl, Br and I;

(h) —OS(O) 2 R 5

wherein R 5 is selected from the group consisting of C 1 -C 24 alkyl groups, C 6 -C 24 aryl groups, C 7 -C 24 alkylaryl groups and C 7 -C 24 arylalkyl groups, groups, aryl groups, alkylaryl groups and arylalkyl groups being optionally substituted;

(i) R 12

wherein R 12 is selected from the group consisting of, unsubstituted or substituted, terminal C 2 -C 24 alkenyl groups, C 3 -C 5 cycloalkenyl groups and C 4 -C 8 alkadienyl groups;

(j) R 13

wherein R 13 is unsubstituted or substituted terminal C 3 -C 24 allenyl group; and

(k) N(R′) 2

wherein R 17 is independently selected from the group consisting of H and C 1 -C 12 alkyl groups;

Z is CH 2 , CF 2 or C(O); or Z is CHOH with the proviso that g is 0, f is 1 and a is 0 or 1;

Y is selected from the group consisting of O, S, N(R 15 ), N(R 15 )C(O), N(R 15 )C(O)N(R 15 ), N(R 15 )C(O)O, OC(O)N(R 15 )S(O) 2 N(R 15 ) and N(R 15 )C(O)N(R 15 )S(O) 2 O, wherein R 15 is independently selected from the group consisting of H, C 1 -C 12 alkyl groups and (U) f -(T) a -A wherein f, a, U, T and A are as defined above; and

R 14 is selected from the group consisting of:

wherein:

a, f, T, A and U are as defined above;

h is 0 or 1; and

W is selected from the group consisting of O, S, NR 15 , NHS(O) 2 O and NHS(O) 2 NR 15 , wherein R 15 is as defined above.

2. The process according to claim 1 , wherein the β-(1,4)-N-acetylgalactosaminyltransferase is Trichoplusia ni β-(1,4)-N-acetylgalactosaminyltransferase.

3. The process according to claim 1 , wherein A is selected from the group consisting of —N 3 , —C(O)CH 3 , —SH, —CH═CH 2 and —CH 2 C≡CH.

4. The process according to claim 1 , wherein the sugar-derivative nucleotide Su(A)-Nuc has formula (15), (16), (17) or (18):

5. The process according to claim 1 , wherein sugar-derivative nucleotide Su(A)-Nuc has formula (19), (20), (21), (22), (23), (24), (25), (26), (65), (66), (67), (68) or (69):

wherein:

R 14 and R 15 are as defined in claim; and

R 16 is independently selected from the group consisting of H and F.

6. The process according to claim 1 , wherein the sugar-derivative nucleotide Su(A)-Nuc has formula (27):

7. The process according to claim 1 , wherein the nucleotide in sugar-derivative nucleotide Su(A)-Nuc is uridine diphosphate (UDP).

8. The process according to claim 1 , wherein the glycan, comprising a terminal GlcNAc moiety, has formula (1), (9), (10) or (11):

9. The process according to claim 1 , wherein the glycoprotein comprising a glycan, comprising a terminal GlcNAc moiety, is according to B formula (7) or (8b):

wherein:

b, d, e and G are as defined in claim 1 ;

y is independently an integer in the range of 1 to 24; and

Pr is a protein.

10. The process according to claim 1 , wherein the glycoprotein, comprising a glycan comprising a terminal GlcNAc moiety, is an antibody.

11. The process according to claim 1 , wherein R 14 is —NHC(O)Me.