SYSTEM AND METHOD FOR pH FORMULATIONS
A system and method for creating a buffer solution having a desired pH value is disclosed. The method uses two known buffer solutions, each with predetermined pH values, and determines a mathematical relationship which defines the amount of each known buffer solution needed to create the buffer solution with the desired pH. This method can then be used to create one or more denaturation graphs, which demonstrate the stability of a protein at a given pH level.
1 . A method of preparing a buffer of a desired pH using a first solution of said buffer having a first pH and a second solution of said buffer having a second pH, comprising:
preparing a plurality of formulations, each having a different fraction of said first solution;
determining the pH of each of said formulations;
recording each of said fractions and said determined pH values;
determining a non-linear best fit mathematical relationship between said fraction and said pH;
entering desired pH value into said non-linear mathematical relationship to determine a fraction of said first solution to be used to achieve said desired pH; and
preparing said buffer of said desired pH using said determined fraction of said first solution.
2 . The method of claim 1 , wherein said first solution is a basic solution and said second solution is an acidic solution.
3 . The method of claim 1 , wherein said non-linear mathematical relationship comprises a cubic relationship.
4 . The method of claim 1 , wherein said non-linear mathematical relationship comprises a fourth order relationship.
5 . The method of claim 1 , wherein said non-linear mathematical relationship comprises a fifth order relationship.
6 . A method of creating a denaturizing graph for a buffer at a desired pH, comprising:
providing a first solution with a first pH and a predetermined minimum amount of chemical denaturant; a second solution having said first pH and a predetermined maximum amount of a chemical denaturant; a third solution having a second pH and a predetermined minimum amount of chemical denaturant; and a fourth solution having said second pH and said predetermined maximum amount of said chemical denaturant;
using the method of claim 1 to combine said first solution and said third solution into a fifth solution having said desired pH and a predetermined minimum amount of chemical denaturant;
using the method of claim 1 to combine said second solution and said fourth solution into a sixth solution having said desired pH and said maximum amount of chemical denaturant;
mixing said fifth and sixth solution in a plurality of samples, each having said fifth and sixth solution in different proportions to create solutions having different molarities of said chemical denaturant;
measuring an observable property of each of said plurality of samples; and
creating a denaturation graph based on said observable property and said molarity.
7 . The method of claim 6 , wherein said observable property comprises fluorescence.
8 . The method of claim 6 , wherein said fifth solution and sixth solution are created in respective containers.
9 . The method of claim 6 , wherein said observable property comprises fluorescence.
10 . The method of claim 6 , wherein said chemical denaturant comprises urea.
11 . The method of claim 6 , wherein said predetermined minimum amount of denaturant comprises no denaturant.
12 . An apparatus for generating a denaturation graph for a buffer at a desired pH, comprising:
a first well comprising a first solution with a first pH and a predetermined minimum amount of chemical denaturant;
a second well comprising a second solution with a first pH and a predetermined maximum amount of chemical denaturant;
a third well comprising a third solution with a second pH and a predetermined minimum amount of chemical denaturant;
a fourth well comprising a fourth solution with a second pH and a predetermined maximum amount of chemical denaturant;
a storage element, comprising a set of instructions adapted to:
use the method of claim 1 to combine said first solution and said third solution into a fifth solution having said desired pH and a predetermined minimum amount of chemical denaturant; and
use the method of claim 1 to combine said second solution and said fourth solution into a sixth solution having said desired pH and said maximum amount of chemical denaturant;
wherein said storage element further comprises said non-linear best fit mathematical relationship;
a controller adapted to execute said instructions; and
a pump or siphon system to draw said first solution from said first well, said second solution from said second well, said third solution from said third well, and said fourth solution from said fourth well in exact quantities and combine said solutions.
13 . The apparatus of claim 12 , wherein said non-linear best fit mathematical relationship comprises a cubic relationship.
14 . The apparatus of claim 12 , further comprising instructions adapted to:
mix said fifth and sixth solution in a plurality of samples, each having said fifth and sixth solution in different proportions to create solutions having different molarities of said chemical denaturant;
measure an observable property of each of said plurality of samples; and
create a denaturation graph based on said observable property and said molarity.
15 . The apparatus of claim 14 , further comprising a detector to determine said observable property.
16 . The apparatus of claim 14 , wherein said observable property comprises fluorescence.
17 . The apparatus of claim 12 , further comprises one or more cannulas to draw said solutions from said wells.
18 . The apparatus of claim 17 , further comprises an actuator to move said cannulas to each of said wells.
19 . The apparatus of claim 14 , wherein said chemical denaturant comprises urea.