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Biological Sciences

Nanoscale Proteomics

Contact: Dick Smith

Proteomics-based biomedical research is increasingly looking for ways to analyze proteins from extremely small amounts of sample, such as from a thousand human blood cells. The sensitivity and dynamic range of proteomics analyses are particularly important when absolute sample size is constrained. To address these needs, we recently developed nano-separations and analysis capabilities that enable protein identifications from nanogram-sized complex biological samples (Shen et al. 2003, Shen et al. 2004). These capabilities include an online micro solid-phase extraction device to maximize peptide recovery and a 15-μm i.d. packed capillary LC for high separation efficiency (20 nL/min, 10,000 psi) that is coupled to an FTICR (using a 5-μm ESI emitter). This platform was recently used to identify proteins from samples that ranged from 50 ng down to 0.50 pg total protein (orders of magnitude smaller than the total amount of proteins predicted for 1000 blood cells).

References

Shen Y, Moore RJ, Zhao R, Blonder J, Auberry DL, Masselon C, Paša-Tolic L, Hixson KK, Auberry KJ, Smith RD. 2003. "High-efficiency on-line SPE voupling to 15-150 mm I.D. column LC for proteomic analysis." Analytical Chemistry 75:3596-3605.

Shen Y, Tolic N, Masselon C, Paša-Tolic L, Camp DGI, Hixson KK, Zhao R, Anderson GA, Smith RD. 2004. "Ultrasensitive proteomics using high-efficiency on-line micro-SPE-NanoLC-NanoESI MS and MS/MS." Analytical Chemistry 76:144-154.

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