Class I MHC Tetramer Preparation: Overview

Production of MHC Tetramers can be broken down into a series of discrete steps, each described on their on page of this website:

Time considerations

Step 1 involves normal molecular biology manipulations such as PCR amplication of the MHC gene fragment, cloning, and sequencing. It can be easily done in a week. There is no important temporal connection between this step and the other steps. Only the general features of the vectors are described here; the details on cloning and sequencing are commonplace and can be found in common molecular biology manuals.
Steps 2 and 3 are connected and can be performed in 2 days. The autoinduction cultures are grown overnight, and collected in the morning. The inclusion bodies are processed that same day.
There is no need or rationale for freezing of E. coli pellets; on the other hand, freezing of the pellets probably doesn't do any harm.
Step 4 can take 2-5 days. As far as I know, the MHC and the BSP tail are stable in folding buffer, and it is not crucial to proceed quickly to the subsequent steps.
Once you begin step 5, it is important to proceed through the completion of step 8 as quickly as possible.
While the globular domains of an MHC molecule are relatively stable and resistant to proteases which might contaminate the preps, the BSP tag is a linear, unstructured entity that is highly susceptible to proteolysis. Rapid completion of the purification through storage at -80 °C will minimize cleavage reactions that inhibit tetramer formation.
The samples for step 9 can be stored at -80 °C prior to assay, so it is not crucial to perform this step immediately. However, there is no reason to not perform this step within a week of the completion of the purification of the tetramer.
Step 10 takes about two hours. Once the monomers are bound to streptavidin, the BSP seems to be more resistant to proteolysis, perhaps because the proteases can't get near the BSP without bumping into the streptavidin.