The 1990 ABRF Amino Acid Analysis study was designed to appraise accuracy and precision of analysis methodology for samples in the presence of relatively high amounts of buffer salts and for samples immobilized on PVDF membranes. Two samples (ABRF-9OAAA2 and ABRF-9OAAA3) plus a control (ABRF-9OAAA1) were distributed to member facilities in mid-March and 37 laboratories returned data by June 1990. Facilities were asked to hydrolyze in triplicate and analyze 90AAA1 (4 ug protein) and 90AAA2 (4 ug protein + 600 ug Tris salt). Facilities were also asked to hydrolyze and analyze three pieces of PVDF with 0.9 ug protein.
As expected, the best results were obtained with the salt-free control sample for which an average accuracy of about 84% was obtained compared with about 73% average accuracy for the protein with Tris buffer. For both 90AAA1 and 90AAA2, about one-third of the participating facilities achieved 90% or greater accuracy. Based upon the results, the protein immobilized on PVDF presented a much greater challenge to available methodology. Overall accuracy (about 62%) and recovery (about 68%) from the PVDF samples were significantly inferior to that obtained with the other two samples.
The results from this study will be detailed at the ABRF Satellite Meeting. Methods used for obtaining the best results from the salt-containing and PVDF immobilized samples will be highlighted. A manuscript describing this study is planned for the volume associated with the Protein Society Meeting which will be edited by Joseph Villafranca.
Daniel Strydom at Brigham and Women's Hospital in Boston has accepted a spot on the 1991 AAA subcommittee, replacing Lowell Ericsson. A new chair remains to be selected. Daniel is well known for practical amino acid analysis and we all look forward to his help with next year's efforts.
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Created: 10th August 1995
Last modified: 10th August 1995
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