High level insect expression of rage proteins

A method for high level expression of recombinant human Receptor for Advanced Glycated Endproducts (RAGE) comprising:

• (a) preparing recombinant virus comprising a nucleotide sequence encoding RAGE or a fragment thereof subcloned into the Autographa californica nuclear polyhedrosis virus;
• (b) preparing a high-titer stock of the recombinant virus;
• (c) initiating cultures of insect cells at an initial density of less than 0.5×106 cells per ml;
• (d) growing the insect cells until the cell density comprises 1 to 20×106 cells per ml;
• (e) adding virus from step (b) at a MOI of less than 30 to the insect cells from step (d) for large scale expression;
• and
• (f) incubating the infected culture at about 26-28° C. under conditions such that a predetermined level of RAGE or a fragment thereof is produced.

A method for high level expression of recombinant human Receptor for Advanced Glycated Endproducts (RAGE) comprising:

• (a) preparing recombinant virus comprising a nucleotide sequence encoding RAGE or a fragment thereof subcloned into the Autographa californica nuclear polyhedrosis virus;
• (b) infecting insect cells at a multiplicity of infection (MOI) of about 0.1 to 0.2;
• (c) incubating the insect cell culture at a temperature ranging from 26-28° C. for 3-7 days to prepare high titer virus stock;
• (d) titering the virus to determine MOI;
• (e) initiating cultures of insect cells at an initial density of about 2.5×105 cells per ml;
• (f) growing the insect cells such that the growth rate comprises a doubling time of about 18-26 hours and the cells comprise a viability of greater than 90% until the cell density comprises 1.5 to 2.5×106 cells per ml;
• (g) adding virus (from step (d)) at a MOI of 0.1 to 10 to the insect cells;
• and
• (h) incubating the infected culture at about 26-28° C. for a predetermined time or until cloudy.