Isolated laminin-421
The present disclosure provides isolated laminin-421, methods for making recombinant laminin-421, and host cells that express recombinant laminin-421. The present disclosure also provides nucleic acid sequences encoding full length human laminin β2 chain, expression vectors and host cells thereof.
1. An isolated recombinant laminin-421 produced by a method comprising:
providing host cells that express recombinant laminin-421, wherein the recombinant laminin-421 comprises:
a first chain comprising a polypeptide with 100% identity to a polypeptide sequence of SEQ ID NO: 1,
a second chain comprising a polypeptide with % identity to a polypeptide sequence of SEQ ID NO: 2, and
a third chain comprising a polypeptide with 100% identity to a polypeptide sequence of SEQ ID NO: 3,
wherein the first, second, and third chains assemble to form a recombinant trimeric laminin-421 structure;
growing the host cells in a cell culture medium under conditions to stimulate expression of the recombinant laminin-421 chains;
passing the cell culture medium through a column, wherein the column contains a compound that binds to the recombinant laminin-421;
washing the column to remove unbound materials; and
eluting the bound recombinant laminin-421 from the column.
2. A pharmaceutical composition, comprising: a) the isolated recombinant laminin-421 of claim 1 ; and b) a pharmaceutically acceptable carrier.
3. An isolated recombinant laminin-421, comprising:
a first chain comprising a polypeptide with 100% identity to a polypeptide sequence of SEQ ID NO: 1;
a second chain comprising a polypeptide with 100% identity to a polypeptide sequence of SEQ ID NO: 2; and
a third chain comprising a polypeptide with 100% identity to a polypeptide sequence of SEQ ID NO: 3;
wherein the first, second, and third chains assemble to form a recombinant trimeric laminin-421 structure.
4. A pharmaceutical composition comprising: a) the isolated recombinant laminin-421 of claim 3 ; and b) a pharmaceutically acceptable carrier.