The entire expression workflow can be tracked in the StackWave Affinity LIMS
Triage
Triage is the first step of the protein production workflow. On this page, users decide whether the requested protein already exists in inventory and thus does not require expression, or if the request should be assigned for expression and which user to assign the task to.
To assign a task for expression, select one or more tasks from the available list and click Assign for Expression, then select a user to assign to. If task emails are enabled, users with assigned tasks will be notified by email at the configured cadence (daily or weekly).
Requests for proteins which are already in inventory and thus do not require expression can be fulfilled here without having to continue through the rest of the workflow. Affinity will automatically check each lot of a tasks' requested protein, aggregate its available amount, and display that on the Triage tab. Users can compare the amount requested with the amount available to determine if Fulfillment from Inventory is possible.
Select the appropriate tasks and click Fulfill from Inventory. This will open the fulfillment modal, which summarizes the details of the request and lists each location in inventory where this protein can be found along with its label, volume, concentration, and amount in mg. The system will automatically select lots from inventory (oldest and largest containers given priority) until the requested amount is fulfilled, however the user is able to change this selection as they see fit.
Affinity will not enforce that the amount checked equals the amount requested, if manual alterations to the selection are made. Once the desired inventory items are selected, click Fulfill in the lower right of the window. Selected lots will be marked for deletion, the requesting user will be notified via email that a task has been completed, and the task will be marked as complete on the Protein Requests page.
Transfect
Transfection is the second step of the protein production workflow. Tasks which have been triaged will appear in the transfect tab as long as there are vectors registered in the system which can be used to express the requested protein. If not, appropriate vector construction tasks will be created in Vector Construction which must first be satisfied before expression.
Tasks can be re-assigned, postponed, or cancelled using the buttons at the top of the form.
To continue with expression, select one or more tasks and click the Express button.
The resulting form allows the user to specify the cell line, plasmids, transfection date, and transfection volume for the protein being expressed.
If the requested entity is a multispecific antibody, the plasmids field will require as many selections as there are chains on the multispecific.
Once the Express button in the lower right is clicked, the proteins will be moved to the next step, Purify.
Purify
Purify is the third tab in the protein production workflow, and allows users to record a series of purification methods and their associated dates, review the history of purification, as well as submit and review SEC Purity data.
To load purification data, select one or more tasks and click Purify.
The resulting form allows for selection of a purification date and method. Submitting this form will not send the task to the next step of the workflow, as its possible to perform purification multiple times before being ready to continue. After submitting at least one purification date/method, the appropriate fields will be populated on the Purify index page.
The Purification Date and Method will show the most recent combination loaded, but the History button can be clicked in the Purification Status form for a full review of purification on the given protein.
SEC Purity Results can be loaded and subsequently reviewed on the Purify index page.
Once a protein is ready to progress, select it on the left and click Complete.
Characterize
Characterization is an optional step of the expression workflow, in which users can upload assay data against the samples associated with the requested proteins to ensure they meet quality control standards. To do so, select one or more tasks on the index page and click Characterize.
The resulting form allows for submission of the yield and concentration, endotoxin (EU/mL), a SEC (% Purity) value, or a SEC Purity Image. Once characterized, the task will be removed from the Characterize index page. Characterization does not need to occur in order to aliquot.
Aliquot
The final step of the protein production workflow is Aliquot. In this step, users specify the amount of the requested protein created and where the resulting material will be sent (i.e. to the requestor or to a specific inventory location).
Select one or more tasks and click Aliquot. In the resulting form, specify either Tube or Vial, select a destination in inventory, and submit the volume, concentration, and number of tubes for inventory and requestor. The final two columns, Amount Requested and Amount Fulfilled, will be system generated.
The volume and concentration are used in combination with the number of tubes for requestor to determine the amount fulfilled. Affinity does not require that the user specify an amount fulfilled which matches the amount requested - these values are provided only for reference. After submission of the form, the task is removed from the aliquot list, marked as complete on the Protein Requests page, and two sets of inventory containers are created - one which is deleted immediately (these are the tubes/vials going to the requestor, but are kept in the system for reference), and another set of tubes/vials depending on what was specified in the “# of Tubes for Inventory” column. The requestor of the proteins will be notified via email that the task is complete.
Lot Review
Lot Review gathers all data which has been uploaded against samples of requested proteins during the protein production workflow process, and presents them in a single form for review purposes. The user is able to mark specific lots as Do Not Use or to Flag them.
Users can apply a filter at the top of the form to show all lots, or only those lots which have been flagged or marked as Do Not Use.
