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Direct identification of on-bead peptides using surface-enhanced Raman spectroscopic barcoding system for high-throughput bioanalysis.

Kang H, Jeong S, Koh Y, Geun Cha M, Yang JK, Kyeong S, Kim J, Kwak SY, Chang HJ, Lee H, Jeong C, Kim JH, Jun BH, Kim YK, Hong Jeong D, Lee YS - Sci Rep (2015)

Bottom Line: The 44 kinds of SERS IDs were able to generate simple codes and could possibly generate more than one million kinds of codes by incorporating combinations of different SERS IDs.The SERS ID encoding based screening platform can identify the peptide ligand on the bead and also quantify its binding affinity for specific protein.We believe that our SERS barcoding technology is a promising method in the screening of one-bead-one-compound (OBOC) libraries for drug discovery.

View Article: PubMed Central - PubMed

Affiliation: Interdisciplinary Program in Nano-Science and Technology, Seoul National University, Seoul 151-744, Republic of Korea.

ABSTRACT
Recently, preparation and screening of compound libraries remain one of the most challenging tasks in drug discovery, biomarker detection, and biomolecular profiling processes. So far, several distinct encoding/decoding methods such as chemical encoding, graphical encoding, and optical encoding have been reported to identify those libraries. In this paper, a simple and efficient surface-enhanced Raman spectroscopic (SERS) barcoding method using highly sensitive SERS nanoparticles (SERS ID) is presented. The 44 kinds of SERS IDs were able to generate simple codes and could possibly generate more than one million kinds of codes by incorporating combinations of different SERS IDs. The barcoding method exhibited high stability and reliability under bioassay conditions. The SERS ID encoding based screening platform can identify the peptide ligand on the bead and also quantify its binding affinity for specific protein. We believe that our SERS barcoding technology is a promising method in the screening of one-bead-one-compound (OBOC) libraries for drug discovery.

No MeSH data available.


Related in: MedlinePlus

Stability and cross-contamination test of SERS nano-identifier (SERS ID)-encoded TentaGel (TG) beads. a) Average number of SERS IDs on the TG beads after treatment with swelling solvents (dichloromethane, DCM), 3% bovine serum albumin (BSA) containing phosphate-buffered saline (PBS, pH 7.0) and 1% Tween 20 containing PBS (pH 7.0). The number was normalized to the total number of SERS IDs on non-treated beads. b) Signal intensities of 488 cm−1 band (4-BBT) and 539 cm−1 band (4-CBT) from SERS spectrum of SERS ID[4BBT]- or SERS ID[4CBT]-encoded TG beads after treatment with 3% BSA containing PBS (pH 7.0), and c) 1% Tween 20 containing PBS (pH 7.0) (The number of measured beads = 6).
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f4: Stability and cross-contamination test of SERS nano-identifier (SERS ID)-encoded TentaGel (TG) beads. a) Average number of SERS IDs on the TG beads after treatment with swelling solvents (dichloromethane, DCM), 3% bovine serum albumin (BSA) containing phosphate-buffered saline (PBS, pH 7.0) and 1% Tween 20 containing PBS (pH 7.0). The number was normalized to the total number of SERS IDs on non-treated beads. b) Signal intensities of 488 cm−1 band (4-BBT) and 539 cm−1 band (4-CBT) from SERS spectrum of SERS ID[4BBT]- or SERS ID[4CBT]-encoded TG beads after treatment with 3% BSA containing PBS (pH 7.0), and c) 1% Tween 20 containing PBS (pH 7.0) (The number of measured beads = 6).

Mentions: The stability of the SERS IDs on the TG beads was assessed in the presence of i) a swelling solvent (dichloromethane), ii) blocking solution (3% bovine serum albumin [BSA] containing phosphate-buffered saline [PBS]), and iii) surfactant containing washing buffer (1% Tween 20 containing PBS). SERS ID-encoded Ac-Gly-TG beads were treated with each solvent for 30 min. After the treatment, the average number of SERS IDs per 9 μm2 area of the TG beads was calculated and normalized to the total number of SERS IDs on non-treated beads, where counts were based on SEM images (representative SEM images are shown in Figure S7). As shown in Fig. 4a, the average number of SERS IDs on the TG beads after treating with each condition did not decrease significantly. To evaluate the SERS ID stability further, cross-contamination tests were performed using the mixture of SERS IDs[4BBT]-encoded TG beads and SERS ID[4CBT]-encoded TG beads. The SERS intensities of 4-BBT and 4-CBT were measured from the mixture of SERS ID[4BBT] attached the TG beads and SERS ID[4CBT] attached the TG beads after treatment with 3 wt% BSA containing PBS solution or with 1 wt% Tween 20 containing PBS solution. As shown in Fig. 4b,c, the SERS intensities from representative peaks (488 cm−1 for 4-BBT and 539 cm−1 for 4-CBT) showed only the original codes. The result indicates that the original encoding for each SERS ID remained intact even after the treatment with the solution containing surfactants, showing no signs of cross-contamination to any significant extent.


Direct identification of on-bead peptides using surface-enhanced Raman spectroscopic barcoding system for high-throughput bioanalysis.

Kang H, Jeong S, Koh Y, Geun Cha M, Yang JK, Kyeong S, Kim J, Kwak SY, Chang HJ, Lee H, Jeong C, Kim JH, Jun BH, Kim YK, Hong Jeong D, Lee YS - Sci Rep (2015)

Stability and cross-contamination test of SERS nano-identifier (SERS ID)-encoded TentaGel (TG) beads. a) Average number of SERS IDs on the TG beads after treatment with swelling solvents (dichloromethane, DCM), 3% bovine serum albumin (BSA) containing phosphate-buffered saline (PBS, pH 7.0) and 1% Tween 20 containing PBS (pH 7.0). The number was normalized to the total number of SERS IDs on non-treated beads. b) Signal intensities of 488 cm−1 band (4-BBT) and 539 cm−1 band (4-CBT) from SERS spectrum of SERS ID[4BBT]- or SERS ID[4CBT]-encoded TG beads after treatment with 3% BSA containing PBS (pH 7.0), and c) 1% Tween 20 containing PBS (pH 7.0) (The number of measured beads = 6).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4446893&req=5

f4: Stability and cross-contamination test of SERS nano-identifier (SERS ID)-encoded TentaGel (TG) beads. a) Average number of SERS IDs on the TG beads after treatment with swelling solvents (dichloromethane, DCM), 3% bovine serum albumin (BSA) containing phosphate-buffered saline (PBS, pH 7.0) and 1% Tween 20 containing PBS (pH 7.0). The number was normalized to the total number of SERS IDs on non-treated beads. b) Signal intensities of 488 cm−1 band (4-BBT) and 539 cm−1 band (4-CBT) from SERS spectrum of SERS ID[4BBT]- or SERS ID[4CBT]-encoded TG beads after treatment with 3% BSA containing PBS (pH 7.0), and c) 1% Tween 20 containing PBS (pH 7.0) (The number of measured beads = 6).
Mentions: The stability of the SERS IDs on the TG beads was assessed in the presence of i) a swelling solvent (dichloromethane), ii) blocking solution (3% bovine serum albumin [BSA] containing phosphate-buffered saline [PBS]), and iii) surfactant containing washing buffer (1% Tween 20 containing PBS). SERS ID-encoded Ac-Gly-TG beads were treated with each solvent for 30 min. After the treatment, the average number of SERS IDs per 9 μm2 area of the TG beads was calculated and normalized to the total number of SERS IDs on non-treated beads, where counts were based on SEM images (representative SEM images are shown in Figure S7). As shown in Fig. 4a, the average number of SERS IDs on the TG beads after treating with each condition did not decrease significantly. To evaluate the SERS ID stability further, cross-contamination tests were performed using the mixture of SERS IDs[4BBT]-encoded TG beads and SERS ID[4CBT]-encoded TG beads. The SERS intensities of 4-BBT and 4-CBT were measured from the mixture of SERS ID[4BBT] attached the TG beads and SERS ID[4CBT] attached the TG beads after treatment with 3 wt% BSA containing PBS solution or with 1 wt% Tween 20 containing PBS solution. As shown in Fig. 4b,c, the SERS intensities from representative peaks (488 cm−1 for 4-BBT and 539 cm−1 for 4-CBT) showed only the original codes. The result indicates that the original encoding for each SERS ID remained intact even after the treatment with the solution containing surfactants, showing no signs of cross-contamination to any significant extent.

Bottom Line: The 44 kinds of SERS IDs were able to generate simple codes and could possibly generate more than one million kinds of codes by incorporating combinations of different SERS IDs.The SERS ID encoding based screening platform can identify the peptide ligand on the bead and also quantify its binding affinity for specific protein.We believe that our SERS barcoding technology is a promising method in the screening of one-bead-one-compound (OBOC) libraries for drug discovery.

View Article: PubMed Central - PubMed

Affiliation: Interdisciplinary Program in Nano-Science and Technology, Seoul National University, Seoul 151-744, Republic of Korea.

ABSTRACT
Recently, preparation and screening of compound libraries remain one of the most challenging tasks in drug discovery, biomarker detection, and biomolecular profiling processes. So far, several distinct encoding/decoding methods such as chemical encoding, graphical encoding, and optical encoding have been reported to identify those libraries. In this paper, a simple and efficient surface-enhanced Raman spectroscopic (SERS) barcoding method using highly sensitive SERS nanoparticles (SERS ID) is presented. The 44 kinds of SERS IDs were able to generate simple codes and could possibly generate more than one million kinds of codes by incorporating combinations of different SERS IDs. The barcoding method exhibited high stability and reliability under bioassay conditions. The SERS ID encoding based screening platform can identify the peptide ligand on the bead and also quantify its binding affinity for specific protein. We believe that our SERS barcoding technology is a promising method in the screening of one-bead-one-compound (OBOC) libraries for drug discovery.

No MeSH data available.


Related in: MedlinePlus