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SpinTip: A Simple, Robust, and Versatile Preanalytical Method for Microscale Suspension Cell Proteomics.

Sebastian E Koschade^1,2,3,4,5, Georg Tascher², Bhavesh S Parmar²

¹Department of Medicine, Hematology/Oncology, University Hospital, Goethe University, 60590 Frankfurt, Germany.

Journal of Proteome Research

|October 14, 2022

Summary

This summary is machine-generated.

Researchers developed a simple SpinTip technique to concentrate scarce suspension cells for microscale proteomics. This method minimizes sample loss, enabling detailed analysis of limited cell samples, including primary acute myeloid leukemia cells.

Keywords:

cell concentration microscale proteomics preanalytical sample loss sample concentration suspension cells volume reduction

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Area of Science:

Biochemistry
Cell Biology
Proteomics

Background:

Preanalytical sample loss and contamination are significant challenges in microscale proteomics, particularly with nonadhering and suspension cells.
Existing workflows are often unsuitable for microscale sample volumes, hindering the analysis of limited cell quantities, such as primary human materials.

Purpose of the Study:

To develop and validate a simple manual batch technique for concentrating scarce suspension cells for microscale applications.
To facilitate preanalytical experimental manipulation and microscale proteomics with limited suspension cell samples.

Main Methods:

A novel technique using pipette tips with heat-sealed orifices (SpinTips) for single-step, 100-fold concentration of suspension cells (down to 5000 cells) by volume reduction.
SpinTips function as microcentrifugation vessels, allowing supernatant aspiration with minimal cell loss, achieving a residual volume of approximately 3 μL.

Main Results:

The SpinTip method effectively concentrates, exchanges medium, washes, and cultures highly limited amounts of suspension cells.
Demonstrated successful application in profiling ex vivo responses of primary acute myeloid leukemia cells to daunorubicin, quantifying 3462 proteins with high repeatability.

Conclusions:

SpinTips offer a versatile and readily incorporated solution for handling scarce suspension cells in microscale proteomics and functional studies.
This technique overcomes preanalytical pitfalls, enabling robust analysis of limited cell samples, including clinical specimens.