Appendix 2 Upc Assay Development - Binding Site Minineph Plus Manual De Usuario

APPENDIX: 2
Below are guidelines for assays using the User Prepared Chemistry mode of the
MININEPH
PLUS
1. Use only MININEPH cuvettes.
2. All sample dilutions must be made using MININEPH Sample Diluent (SN042).
3. Ensure that the analyser is primed with an appropriate on-line buffer. UPC mode
always draws its on-line buffer from line „B‟.
4. Begin with a blank time of 5 seconds and a read time of 600 seconds.
5. The pipette aspiration sequence depends on the chosen assay temperature. For a
low temperature assay, the sequence is:
Aspirate 400L (e.g. Reaction buffer)
Aspirate 40L (e.g. Air gap)
Aspirate 40L (e.g. Reagent)
Dispense and blow out all reagents
For a high temperature assay, the sequence is
Aspirate 40L (e.g. Reagent)
Aspirate 40L (e.g. Air gap)
Aspirate 100L (e.g. Supplementary reagent)
Dispense 480L (All aspirated reagents plus on-board buffer)
6. Perform assays at doubling dilutions of the assay calibration fluid to determine the
range. Begin with a sample volume of 40L.
When the test range has been determined, select an appropriate sample test dilution
for this range.
If a suitable assay range cannot be determined the following parameters may be
adjusted:
(1) Antiserum dilution
(2) Sample dilution or volume
(3) Assay time (blank time or read time).
7. The scatter units for the top point of the curve should be in the range 2000-3000. The
scatter units for the bottom point should be between 100-500 units.
8. Avoid curves that may suffer antigen excess problems. If antigen excess is
suspected repeat the assay using a less dilute sample. If this results in an increase in
the scatter units there is no antigen excess problem.
9. It is preferable to use a curve-fitting program. Errors for such a curve should be within
specified limits (+/- 5% is recommended). If errors fall outside this range, consider
further changes to the assay conditions.
Insert Code: EIN023
UPC ASSAY DEVELOPMENT
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Version: 11 August 2011