UTHSCSA Center for Surface Plasmon Resonance (CSPR)

Sample BIA method file

This is an example of a BIA method file. For your particular method you may need more or less positions for samples or may want to group all the blanks in one row. In this particular example all samples are treated the same way, with a kinject of 150 µL (1800 seconds or 30 minutes of contact time) followed by a 1800 second dissociation time and then a 5 second regeneration step from position r2f3 with extracleanup. The analytes are flowed over all 4 flow cells of the sensor chip at a flow rate of 5 µL per minute. The program loops through the steps s1..s2.........s50. Once the program ends, it tells the instrument to enter standby mode. Each cycle lasts approximately 1 hour and 8 minutes in this example.

A text version is available for download and appropriate modifications.

DEFINE LOOP cycle1
LPARAM %s1 %p1

times 1

s1 r2a1  ! indicates sample 1 located in rack 2 (located on your right) position a1

s2 r2a2  ! generally samples r2a1 - r2a10 are buffer blanks which allows the instrument to "settle down."

s3 r2a3  ! These buffer blanks are hidden in Scrubber.

s4 r2a4

s5 r2a5

s6 r2a6

s7 r2a7

s8 r2a8

s9 r2a9

s10 r2a10

s11 r2b1  ! Lowest concentration of first analyte to flow over the sensor chip

s12 r2b2  ! Next highest concentration of the first analyte and so forth with a total of seven (7) different concentrations.

s13 r2b3

s14 r2b4

s15 r2b5

s16 r2b6  ! Highest concentration of first analyte.

s17 r2b7

s18 r2b8  ! Buffer blank to be used in Scrubbers double referencing system

s19 r2b9  ! Buffer blank to be used in Scrubbers double referencing system

s20 r2b10  ! Buffer blank to be used in Scrubbers double referencing system

s21 r2c1  ! Lowest concentration of replicate of first analyte or lowest concentration of second analyte. Repeat as above.

s22 r2c2

s23 r2c3

s24 r2c4

s25 r2c5

s26 r2c6

s27 r2c7

s28 r2c8  ! Buffer blank to be used in Scrubbers double referencing system

s29 r2c9  ! Buffer blank to be used in Scrubbers double referencing system

s30 r2c10  ! Buffer blank to be used in Scrubbers double referencing system

s31 r2d1  ! Lowest concentration of replicate of first analyte or lowest concentration of third analyte. Repeat as above.

s32 r2d2

s33 r2d3

s34 r2d4

s35 r2d5

s36 r2d6

s37 r2d7

s38 r2d8  ! Buffer blank to be used in Scrubbers double referencing system

s39 r2d9  ! Buffer blank to be used in Scrubbers double referencing system

s40 r2d10  ! Buffer blank to be used in Scrubbers double referencing system

s41 r1a1  ! indicates sample 1 located in rack 1 (located on your left) position a1

s42 r1a2

s43 r1a3

s44 r1a4

s45 r1a5

s46 r1a6

s47 r1a7

s48 r1a8

s49 r1a9

s50 r1a10


END

DEFINE APROG analyse1
PARAM %s1 %p1
flow 5 -f  ! flow rate is set to 5 µL/min
flowpath 1,2,3,4  ! the analyte flows over all 4 flow cells
kinject %p1 150 1800 : ! the 150 refers to the µL of analyte to flow over the sensor chip; the 1800 refers to the number of seconds to allow for dissociation; both "on" and "off" times are 30 minutes
quickinject r2f3 5  ! regeneration with extraclean
extraclean

wash i  ! wash all IFC channels
wash f  ! wash flow cell area

end

MAIN
DETECTION 1,2,3,4

LOOP cycle1 STEP
APROG analyse1 %s1 %p1
ENDLOOP


append standby  ! standby procedure initiated after completion of the program
END