From a lateral flow kit commercial that uses gold particles with antibodies as a visual marker to detect Plasmodium (LDH protein), I am trying to recuperate this protein separating the immune complex formed in the positive line of the test. I have tried some lysis buffer to dissociate the immune complex without denaturing the protein, using HCl-glycine and detergents as NP-40. However, the performance is poor and the positive line remains similar. I am using the visual line as a surrogate marker that the dissociation is not working. My questions are: Is it possible to dissociate the immune complex from the kit in the conditions described above? Is it right to use the loss of colour of the positive line as a control of the immune complex dissociation?
You are of course aware that the amount of protein recuperated this way will be tiny. I would try the methods published for stripping of western-blots (for example, DOI: 10.1002/elps.201200333).
As Engelbert has mentioned, the fraction of total specific protein on the detection line is low. You could try to extract more of the strip and possibly concentrate later by using an immunoaffinity column with covalently immobilized antibodies (e.g. using NHS activated sepharose). If you should need more plasmodium LDH, go directly on such a column. But keep in mind that the acidic or alkalie elution process might inactivate your enzyme, if this should be of concern.
Thank you very much Wolfgang Schechinger and Engelbert Buxbaum for your help. Effectively the performance was being minimal. I think I am going to start directly from the sample and try to isolate the protein using immnoaffinity columns.
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