1. A process for preparing a compound of formula (I-a):
comprising, contacting 5,6-difluoro-N1-(2-fluoro-4-iodophenyl)-3-methoxybenzene-1,2-diamine with a compound of formula (II-a):
wherein
R1 is H or an alcohol protecting group;
R2 is H or an alcohol protecting group; or
R1 and R2 together with the carbon atoms to which they are attached form a cyclic 1,2-diol protecting group.
2. A process for preparing a compound of formula (I-b):
comprising, contacting 5,6-difluoro-N1-(2-fluoro-4-iodophenyl)-3-methoxybenzene-1,2-diamine with a compound of formula (II-b):
wherein
R1 is H or an alcohol protecting group;
R2 is H or an alcohol protecting group; or
R1 and R2 together with the carbon atoms to which they are attached form a cyclic 1,2-diol protecting group.
3. A process of claim 1, wherein R1 and R2 together with the carbon atoms to which they are attached form a cyclic 1,2-diol protecting group.
4. The process of claim 3, wherein the cyclic 1,2-diol protecting group is a 5-membered cyclic 1,2-diol protecting group.
5. The process of claim 4, wherein the cyclic 1,2-diol protecting group is 2,2-dimethyl-1,3-dioxolan-4-yl.
6. A process of claim 2, wherein R1 and R2 together with the carbon atoms to which they are attached form a cyclic 1,2-diol protecting group.
7. The process of claim 6, wherein the cyclic 1,2-diol protecting group is a 5-membered cyclic 1,2-diol protecting group.
8. The process of claim 6, wherein the cyclic 1,2-diol protecting group is 2,2-dimethyl-1,3-dioxolan-4-yl.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A method for identifying the presence of an antigen or an antibody in a sample, the method comprising:
contacting the sample with a complementary capture antibody or a capture antigen in an aqueous solution,
specifically binding the antigen or the antibody to the complementary capture antibody or capture antigen, respectively; and
detecting the presence of the antigen or antibody bound to the complementary capture antibody or capture antigen with a detector antibody that binds to either (a) the antigen, which is bound to the complementary capture antibody, or (b) the antibody, which is bound to the capture antigen, the sample, the capture antibody or capture antigen, the aqueous solution, and the detector antibody forming a matrix, the matrix comprising a first protein that binds to said capture antibody, capture antigen, detector antibody, antigen or antibody in the sample to be detected with a KD value of up to 10\u22127 andor a second protein that binds to said capture antibody, capture antigen, detector antibody, antigen or antibody in the sample to be detected with a KD value of between 10\u22127 and 10\u22129;
said aqueous solution comprising:
a) a buffer to control pH;
b) a compound A selected from the group consisting of polyalkylene glycol, polypropylene glycol, propylene glycol, polyethylene glycol, ethylene glycol, monosaccharides, disaccharides, trisaccharides, saccharose, mannose, trehalose, polyol, glycerol and mixtures thereof; and
c) a non-ionic detergent;
wherein the concentration of compound A is from 2.0-15.0% and the concentration of the non-ionic detergent is from 0.1-1.0%, wherein the concentration of compound A is in terms of % (vv) when compound A is a liquid or the concentration of compound A is in terms of % (wv) when compound A is a solid
wherein the ratio of the non-ionic detergent to the compound A is from 1:15 to 1:25:
wherein the non- ionic detergent is a compound selected from the group consisting of:
a) a substituted phenyl residue having substituents R1 and R2 (R1-Ph- R2), wherein R1 is C1-C9 alkyl group, R2 is a \u2014O\u2014CH2\u2014CH2\u2014O3\u2014H group, wherein \u201ca\u201d is an integer of 5 to 40, wherein R2 in respect to R1 is in para, meta or ortho position,
b)
wherein n, x, y and z together is an integer of 5 to 40, R is a fatty acid residue;
c) Dodecylpoly(ethyleneglycolether)m, wherein m is an integer of 5 to 40;
d) 1-O-n-Octyl-\u03b2-D-glucopyranoside (n-Octylglucoside); and
e) 1-O-n-Dodecyl-\u03b2-D-glucopyranosyl (1-4)alpha-D-glucopyranoside;
wherein said aqueous solution reduces the binding of said first protein or said second protein in the sample to any one or more of the following: complementary capture antibody or capture antigen or detector antibody or the antigen or the antibody in the sample to be detected, thereby reducing interference by the first protein or the second protein with the specific binding of said antigen or antibody to the complementary capture antibody or capture antigen or detector antibody, compared to conducting said specific binding in the absence of said aqueous solution.
2. The method of claim 1, wherein the aqueous solution has an ionic strength of 100 mM to 800 mM.
3. The method of claim 1, wherein the aqueous solution has an ionic strength of 100 mM to 600 mM.
4. The method of claim 1, wherein the aqueous solution has an ionic strength of 100 mM to 500 mM.
5. The method of claim 1, wherein the aqueous solution has an ionic strength of 200 mM to 600 mM.
6. The method of claim 1, wherein the aqueous solution has an ionic strength of 250 mM to 600 mM.
7. The method of claim 1, wherein the solution comprises a salt selected from the group consisting of NaCl, KCl, and NH4Cl.
8. The method of claim 1, wherein said aqueous solution further comprises a blocking protein in an amount effective to immunologically block non-specific antibody binding.
9. The method of claim 8, wherein the blocking protein is selected from the group consisting of bovine serum albumin, ovalbumin, casein, and fetal bovine serum.
10. The method of claim 8, wherein the concentration of the blocking protein is in the range of 0.1 to 2% wv.
11. The method of claim 1, wherein the buffer is selected from the group consisting of Tris (Tris(hydroxymethyl)-aminomethane, Pipes (Piperazine-1,4-bis-2-ethane sulfonic acid), Mes (4-Morpholino ethane sulfonic acid), Hepes (4-(2-hydroxyethyl)-1-piperazine- ethane sulfonic acid), and phosphate buffer.
12. The method of claim 1, wherein the non-ionic detergent is selected from the group consisting of Alkylphenolpoly(ethylene-glycolether)m, wherein m is an integer of 5 to 40; Dodecylpoly-(ethyleneglycolether)m, wherein m=23 (Brij35\xae); Poly(oxyethylene)(20)-sorbitane mono fatty acid ester; Poly(oxyethylene)(20)-sorbitane monooleate (Tween\xae80); Poly(oxyethylene) (20)-sorbitane monolaurate (Tween\xae20); Poly(oxyethylene)(20)-sorbitane monopalmitate (Tween\xae40); Poly(oxyethylene)(20)-sorbitane monostearate); and Octylphenolpoly(ethylene-glycolether)m, wherein m is an integer of 5 to 40.
13. The method of claim 1, wherein the non-ionic detergent is selected from the group consisting of Alkylphenolpoly(ethylene-glycolether)m, wherein m=11 (Nonidet P40\xae); and Octylphenolpoly(ethylene-glycolether)m, wherein m=10 (Triton\xaeX 100).
14. The method of claim 1, wherein the aqueous solution does not contain dithiothreitol.
15. The method of claim 1, wherein the pH is adjusted in the range of 5.6 to 9.6.
16. The method of claim 1, wherein the sample is blood plasma or blood serum.
17. The method of claim 1, wherein the antigen to be identified is C-reactive protein.
18. The method of claim 1, wherein the first protein or the second protein is selected from the group consisting of rheuma factors and hemoglobin.