Efficient and specific removal of albumin from human serum samples.
Patient serum or plasma is frequently monitored for biochemical markers of disease or physiological status. Many of the rapidly evolving technologies of proteome analysis are being used to find additional clinically informative protein markers. The unusually high abundance of albumin in serum can interfere with the resolution and sensitivity of many proteome profiling techniques. We have used monoclonal antibodies against human serum albumin (HSA) to develop an immunoaffinity resin that is effective in the removal of both full-length HSA and many of the HSA fragments present in serum. This resin shows markedly better performance than dye-based resins in terms of both the efficiency and specificity of albumin removal. Immunoglobulins are another class of highly abundant serum protein. When protein G resin is used together with our immunoaffinity resin, Ig proteins and HSA can be removed in a single step. This strategy could be extended to the removal of any protein for which specific antibodies or affinity reagents are available.
Steel
Laura F
LF
Department of Biochemistry and Molecular Pharmacology, Thomas Jefferson University, Jefferson Center for Biomedical Research, Doylestown, Pennsylvania 18901, USA. laura.steel@jefferson.edu
Trotter
Michael G
MG
Nakajima
Pamela B
PB
Mattu
Taj S
TS
Gonye
Gregory
G
Block
Timothy
T
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D001797:Blood Protein Electrophoresis; D004396:Coloring Agents; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011108:Polymers; D040901:Proteomics; D044967:Serum; D012709:Serum Albumin / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014227:Triazines
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D004396:Coloring Agents; C034284:IgG Fc-binding protein, Streptococcus; D007074:Immunoglobulin G; D011108:Polymers; D012709:Serum Albumin; D014227:Triazines; C012842:Cibacron Blue F 3GA