Author: Oude Munnink, Bas B.; Jazaeri Farsani, Seyed Mohammad; Deijs, Martin; Jonkers, Jiri; Verhoeven, Joost T. P.; Ieven, Margareta; Goossens, Herman; de Jong, Menno D.; Berkhout, Ben; Loens, Katherine; Kellam, Paul; Bakker, Margreet; Canuti, Marta; Cotten, Matthew; van der Hoek, Lia
Title: Autologous Antibody Capture to Enrich Immunogenic Viruses for Viral Discovery Document date: 2013_11_4
ID: 0mu4tkui_10
Snippet: Respiratory and fecal samples were centrifuged (10,000 g) and 150 ml of the supernatant was mixed with 50ml Dynabeads protein A and G (1:1, Invitrogen). After 20 minutes incubation, 10 ml of autologous convalescent serum of the patient was added to the mixture (Table 1) . After a 20 minutes incubation with continuous shaking at room temperature, samples were washed six times with PBS using a magnetic particle concentrator. Universal transport med.....
Document: Respiratory and fecal samples were centrifuged (10,000 g) and 150 ml of the supernatant was mixed with 50ml Dynabeads protein A and G (1:1, Invitrogen). After 20 minutes incubation, 10 ml of autologous convalescent serum of the patient was added to the mixture (Table 1) . After a 20 minutes incubation with continuous shaking at room temperature, samples were washed six times with PBS using a magnetic particle concentrator. Universal transport medium with TURBO TM DNase (2U/ml, Ambion) was added to the antibody-antigen complex and samples were incubated at 37uC for 30 minutes. The complexes were lysed with Boom-lysis buffer L6 and the lysate was used as input for Boom extraction [14] , followed by VIDISCA-454 sequencing as described below.
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