Interfacing with the Biacore instrument Different ways of interfacing with the instrument Manual run Wizards Method Builder methods 2 1
Manual Run 3 Application Wizards Guides you through each step in the analysis 4 2
Method Builder Method Builder - Concept Designed for maximum flexibility Built in support for evaluation Integrated assay knowledge 6 3
Method Builder Support Provided Small molecules» Solvent correction» Extra wash procedures Affinity in Solution Buffer change Adaptivity» If Then commands 7 Methods 1(2) Assays are built up step by step in Methods» Possible to save, share and re-use methods 3 approaches to creating methods:» Create Methods from wizards» Start with a Biacore Method» Create your own Method 8 4
Methods 2(2) Pre-defined Biacore Methods» LMW screen» LMW kinetics» Kinetics heterogeneous analyte» Affinity in solution» NTA kinetics» L1 liposome capture 9 Workflow Overview General settings Define method Define run settings and start the run Assay Steps Cycle Types Sample & Assay Setup Verification Setup Run Detection Enter variables Cycle Run List System preparations Rack Positions Save Method (.method) Save Run (.blr) 10 5
Define Method 1(3) Overview General settings Assay Steps Cycle Types Sample & Assay Setup Verification Setup Run Detection Enter variables Cycle Run List System preparations Rack Positions Save Method (.method) Save Run (.blr) 11 Define Method 2(3) Assay Steps» Define the purpose of assay components (e.g. start-up, sample, control sample)» Assay steps can be repeated or recurrent» Recurrent for e.g. control samples every 10 th sample Cycle Types» Define the injection sequence for a cycle using commands» Set parameters for each injection» Parameters can be defined as variables Connect Cycle Type to Assay Step 12 6
Define Method 3(3) Overview General settings Assay Steps Cycle Types Sample & Assay Setup Verification Setup Run Detection Enter variables Cycle Run List System preparations Variable definition In method At run time Some in method, others at run time Rack Positions Save Method (.method) Save Run (.blr) 13 Setup Run Overview General settings Assay Steps Cycle Types Sample & Assay Setup Verification Setup Run Detection Enter variables Cycle Run List System preparations Define run settings and start the run Rack Positions Save Method (.method) Save Run (.blr) 14 7
More about Method Builder Overview General settings Assay Steps Cycle Types Sample & Assay Setup Verification Setup Run Detection Enter variables Cycle Run List System preparations Create method from Kinetics wizard template Rack Positions 15 Overview Provides a summary of the method (settings cannot be changed here) Left panel shows assay steps Right panel shows settings and cycle type definition for the selected assay step 16 8
General Settings At start: Concentration unit At start: Data collection rate At start: Sample compartment temperature After run: Analysis temperature Detection (single, dual or multi)» Affects which flow paths will be available for each injection command 17 Assay Steps Determines the main structure of the method Steps at the top level are executed in the order given Nested steps (marked with an arrow) are executed within the step they are placed 18 9
Cycle Types Injection parameters are set (contact time, type of injection, etc) Report points may be added or altered Variables may be added or altered For example, one cycle type can include the commands Sample1 and Regeneration1 19 Connect Cycle Type to Assay Step Performed in Assay Steps 20 10
Sample & Assay Setup Determine whether variables are defined in method, at run time, or a mixture of these 21 Verification Checks that the method is correctly and completely defined A method that fails verification can be saved but not run Verification may fail because parameters are missing or the method construction is invalid 22 11
Setup Run - Detection Set the flow path for the detection Only choose flow path consistent with Detection setting for the method 23 Setup Run - Variables Assign values to variables that were not defined in method (for each assay step) 24 12
Setup Run Cycle Run List Complete summary of the cycles in the run 25 Setup Run System Preparations Choose preparation steps to be executed before the run Temperature settings can only be defined in method 26 13
Setup Run Rack Positions Displays the sample and reagent positioning 27 Setup Run Prepare Run Protocol Allows you to enter a run protocol with instructions A general protocol is suggested 28 14
Method Builder - Summary Software tool for user-defined methods» Maximum flexibility» Support features not available in wizards» Built-in evaluation support» Biacore Methods provides user-guidance 29 15