I am not an expert in this field, but I am very interested and have researched to find an answer. I received some assistance from tlooto.com for this response. Could you please review the response below to see if it is correct?

To remove immunoglobulins from small volumes of serum, Protein A or Protein G affinity chromatography can be utilized, as they specifically bind to the Fc region of immunoglobulins. Prepare a small column with Protein A or G agarose beads, equilibrate with binding buffer, and load your serum sample. Wash the column to remove unbound proteins and elute the serum without immunoglobulins. Alternatively, consider using immunodepletion kits designed for small volumes, which include ready-to-use beads and simplified protocols, minimizing the loss of target proteins [1][2][3][4][5]. These methods ensure effective removal of immunoglobulins and reduce false positives in western blotting tests.

Reference

[1] Khan, S. R., Burgh, A. C. v. d., Peeters, R., Hagen, P. M. v., Dalm, V. A., & Chaker, L. (2021). Determinants of Serum Immunoglobulin Levels: A Systematic Review and Meta-Analysis. Frontiers in Immunology, 12.

[2] Peppas, I., George, G., Sollie, S., Josephs, D., Hammar, N., Walldius, G., Karagiannis, S., & Hemelrijck, M. v. (2020). Association of Serum Immunoglobulin Levels with Solid Cancer: A Systematic Review and Meta-analysis. Cancer Epidemiology, Biomarkers & Prevention, 29, 527 - 538.

[3] Webster, R., Fahie-Wilson, M., Barker, P., Chatterjee, V., & Halsall, D. (2010). Immunoglobulin interference in serum follicle-stimulating hormone assays: autoimmune and heterophilic antibody interference. Annals of Clinical Biochemistry, 47, 386 - 389.

[4] Slungaard, A., & Smith, M. J. (2009). Serum immunoglobulin levels in chronic lymphatic leukaemia.. Scandinavian journal of haematology, 12 2, 112-6 .

[5] Ojuawo, A., Milla, P., & Lindley, K. (1998). Serum immunoglobulin and immunoglobulin G subclasses in children with allergic colitis.. West African journal of medicine, 17 3, 206-9 .

I am not an expert but Protein A/G/L Affinity Chromatography. Protein A, G, or L bind specifically to the Fc region of immunoglobulins, which allows you to remove them from the serum.

Follow these steps

• Incubate your serum with Protein A/G/L agarose beads.
• Allow the immunoglobulins to bind to the beads.
• Centrifuge to pellet the beads, separating them from the supernatant.
• Collect the supernatant, which should be depleted of immunoglobulins.

Alternatively, You can use an Immunodepletion Kit thats commercially available, the process is more or less the same as the above but its easier and faster than doing the whole thing manually

for small volume of samples, using protein L conjugated beads for immunopreciptation is the best way to remove Igs from the blood samples. after IP, take the top for your western blot..