Affinity Chromatography of F(ab′)2 Fragments of IgG Autoantibodies on IgM Coupled to Sepharose
| . | Loaded Antibodies . | Eluted Antibodies . | Eluted Specific Activity/Loaded Specific Activity . | ||
|---|---|---|---|---|---|
| . | Amount (mg) . | Specific Activity (OD/μg) . | Amount (μg) . | Specific Activity (OD/μg) . | . |
| IVIgM-Sepharose | |||||
| F(ab′)2 of Hashimoto IgG | 2.4 | 0.351 | 1.5 | 45 | 128 |
| F(ab′)2 of anti-FVIII IgG | 1.8 | 0.120 | 4.0 | 2 | 16.67 |
| Control IgM-Sepharose | |||||
| F(ab′)2 of Hashimoto IgG | 0.8 | 0.403 | 0.330 | 0.694 | 1.72 |
| F(ab′)2 of anti-FVIII IgG | 0.374 | 0.170 | 0.144 | 0.015 | 0.08 |
| . | Loaded Antibodies . | Eluted Antibodies . | Eluted Specific Activity/Loaded Specific Activity . | ||
|---|---|---|---|---|---|
| . | Amount (mg) . | Specific Activity (OD/μg) . | Amount (μg) . | Specific Activity (OD/μg) . | . |
| IVIgM-Sepharose | |||||
| F(ab′)2 of Hashimoto IgG | 2.4 | 0.351 | 1.5 | 45 | 128 |
| F(ab′)2 of anti-FVIII IgG | 1.8 | 0.120 | 4.0 | 2 | 16.67 |
| Control IgM-Sepharose | |||||
| F(ab′)2 of Hashimoto IgG | 0.8 | 0.403 | 0.330 | 0.694 | 1.72 |
| F(ab′)2 of anti-FVIII IgG | 0.374 | 0.170 | 0.144 | 0.015 | 0.08 |
F(ab′)2 fragments of IgG purified from the serum of a patient with Hashimoto's thyroiditis and a patient with anti-F.VIII autoimmune disease were loaded on IVIgM-Sepharose and on IgM purified from the plasma of a patient with Waldenström's macroglobulinemia, coupled to Sepharose, for 2 hours at room temperature. The columns were eluted at acid pH. The amount of antibody and specific anti-TG and anti-FVIII autoantibody activity were then assessed in the loaded material and in the acid-eluted fractions of the columns.