BODIPY-based dyes for digital spatial proteomics
AssigneeTrustees of Boston University
InventorsAaron Beaty Beeler, Andrew Emili, Jason Michael Lenihan, Nathaniel Ryan Hendrick, Ichun Anderson Chen
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Abstract
The disclosure provides probes comprising dipyrromethane-BF2 derivatives which exhibits different fluorescent spectral properties when conjugated to the amino acids, compositions and kits comprising same. The disclosure also provides methods for detecting/identifying amino acids and sequencing polypeptide molecules by conjugating a dipyrromethane-BF2 derivative which exhibits different fluorescent spectral properties when conjugated to the amino acids.
Claims28
A method of identifying amino acids in a plurality of amino acids, the method comprising:
- (a) conjugating a probe to amino acids in a plurality of amino acids, wherein the amino acids in the plurality of amino acids are different, wherein the probe comprises a dipyrromethane-BF2 derivative, and wherein the dipyrromethane-BF2 derivative comprises a labile or reactive group and exhibits different fluorescent spectral properties when conjugated to the amino acids in the plurality of the amino acids, wherein each amino acid in the plurality of amino acids is at the N- or C-terminal of a polypeptide;
- (b) detecting one or more fluorescent spectral properties of the probe conjugated to the different amino acids;
- and
- (c) identifying the amino acids in the plurality of the amino acids by comparing the fluorescent spectral properties of the conjugated probe to a plurality of reference fluorescent spectral properties, wherein each reference fluorescent spectral property is representative of the probe conjugated to a different amino acid.
A method of sequencing a polypeptide, the method comprises:
- (a) conjugating a probe to a terminal amino acid of a polypeptide, wherein the probe comprises a dipyrromethane-BF2 derivative, and wherein the dipyrromethane-BF2 derivative comprises a labile or reactive group and exhibits different fluorescent spectral properties when conjugated to different amino acids;
- (b) detecting one or more fluorescent spectral properties of the probe conjugated to the amino acid;
- and
- (c) identifying the amino acid by comparing the fluorescent spectral properties of the conjugated probe to a plurality of reference fluorescent spectral properties, wherein each reference fluorescent spectral property is representative of the probe conjugated to a different amino acid;
- (d) cleaving the conjugated amino acid from the polypeptide;
- and
- (e) sequentially repeating steps (a) to (d) one or more times to determine the sequence of at least a portion of the polypeptide.
Dependencies are inferred from the claim text.