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Neutralizing Antibody (NAb) Assays can be of several different types:

  • Low-throughput assays typically have 96 wells in a single plate, prepared with five specimens in eight dilutions of two replicates each. Low-throughput samples are diluted within a single plate.
  • High-throughput assays typically have 384 wells per plate and may consist of up to eight plates. High-throughput assays have two options for dilution:
    • Cross Plate Dilution: Each well on a given plate has the same dilution level; dilutions occur across plates.
    • Single Plate Dilution: Dilutions occur within a single plate.
  • Multi-Virus NAb assays include multiple viruses within a single plate and may be either low- or high-throughput and either single- or cross-plate dilution.
The specific file format generated by your plate reader will determine how you configure your assay design to properly parse the data and metadata it contains.

Low-Throughput NAb Assay Formats

LabKey's low-throughput NAb assay supports a few different formats. Files containing these formats may be of any type that the TabLoader can parse: i.e. Excel, tsv, csv, txt.

  • format1.xls has the plate data in a specific location on the second sheet of the workbook. The 96 wells of plate data must be in exactly the same location every time, spanning cells A7-L14.
  • SpectraMax.csv contains plate data identified by a "Plate:" cell header.
  • format2.xls follows a more general format. It can have the plate data on any sheet of the workbook, but the rows and columns must be labeled with 1-12 and A-H.
  • format3.tsv is the most general format. It is a file that just contains the plate data without row or column headers.
For all formats, only the plate data is read from the file. All other content, including worksheet naming, is ignored.

Metadata Input Format

For low-throughput NAb assays, sample and virus metadata input may be done manually via form input at assay import time, or may be uploaded as a separate file to simplify the import process and reduce user error. For example, if you are using a FileMaker database you can export the information and upload it directly. The file upload option supports Excel, tsv, csv, or txt file types.

High-Throughput NAb Assay Formats - Metadata Upload

In order to support different plate readers for high-throughput 384-well NAb assays, LabKey tools support two methods of uploading metadata. Some instruments output metadata and run data in separate spreadsheets, others generate a combined file which contains both. As part of new LabKey Assay design, you select a Metadata Input Format of either:

  • File Upload (metadata only): upload the metadata in a separate file.
  • Combined File Upload (metadata & run data): upload both in a single file.
If you are importing a run where the metadata is uploaded in a separate file, you can download a template from the run properties page by clicking Download Template.

Multi-Virus NAb Assay Format

When working with multiple viruses on a single plate, the assay design can be configured to specify multiple viruses and control well groups within a single plate design so that a run may be uploaded at once but results viewed and graphed on a per-virus basis. See Work with Multiple Viruses per Plate for details.

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