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B lymphocyte stimulus factor inhibiting peptide and its screening and preparing method

A technology of B lymphocytes and screening methods, applied in the field of small peptides and their screening and preparation, can solve the problems that cannot be used in in vivo research, and achieve the effect of simple artificial synthesis and purification, and easy large-scale production

Inactive Publication Date: 2006-06-07
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, phage-presenting small peptides cannot be used for in vivo research due to the presence of phage components. That is to say, only the artificial synthesis of the screened phage-presenting small peptides has real development and application value.

Method used

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  • B lymphocyte stimulus factor inhibiting peptide and its screening and preparing method
  • B lymphocyte stimulus factor inhibiting peptide and its screening and preparing method
  • B lymphocyte stimulus factor inhibiting peptide and its screening and preparing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] 1. Screening of phage conformational peptide library with BLyS as target molecule

[0074] 1. Affinity screening of phage peptide library

[0075] ① Add 1.5ml 100mg / L rhBLyS 112-285 Add to all-plastic petri dish, 4 ℃ overnight. The protein solution was discarded, and 5g / LBSA (bovine serum albumin) was added to block at room temperature for 1h.

[0076] ② After washing 8 times with 1 mL / L TBS / Tween20, add 10 μL human phage conformational 7-peptide library (containing 2×10 11 Virus particles), human phage conformation type 7 peptide library including host bacteria ER2738, purchased from NewEngland Biolabs, and adsorbed at room temperature for 55min.

[0077] Wherein the formula of TBS buffer solution is Tris-HCl 50mmol / L, NaCl150mM / L.

[0078] ③ After washing 7 times with 1 mL / L TBS / Tween20, add 1 mL of 0.2 mol / L HCl-Gly (glycine) elution buffer with a pH value of 2.2 and 1 g / L BSA, dissociate at room temperature for 7 minutes, and then add 150 μL of pH value 9.1 1mo...

Embodiment 2

[0117] 1. Screening of phage conformational peptide library with BLyS as target molecule

[0118] 1. Affinity screening of phage peptide library

[0119] ①Add 120mg / L rhBLyS 112-285 Add to polystyrene container, 4 ℃ overnight. The protein solution was discarded, and 5g / LBSA (bovine serum albumin) was added to block at room temperature for 1h.

[0120] ② After washing 9 times with 1 mL / L TBS / Tween20, add 10 μL human phage conformational 7-peptide library (containing 2×10 11 Virus particles), human phage conformation type 7-peptide library including host bacteria ER2738, purchased from NewEngland Biolabs, and adsorbed at room temperature for 60min.

[0121] Wherein the formula of TBS buffer solution is Tris-HCl 50mmol / L, NaCl150mM / L.

[0122] ③ After washing 6 times with 1 mL / L TBS / Tween20, add 1 mL of 0.2 mol / L HCl-Gly (glycine) elution buffer with a pH value of 2.2 and 1 g / L BSA, dissociate at room temperature for 8 min, and then add 150 μL pH value 9.1 1mol / L Tris-HCl ne...

Embodiment 3

[0128] 1. Screening of phage conformational peptide library with BLyS as target molecule

[0129] 1. Affinity screening of phage peptide library

[0130] ①Add 150mg / L rhBLyS 112-285 Add to polystyrene container, 4 ℃ overnight. The protein solution was discarded, and 5g / LBSA (bovine serum albumin) was added to block at room temperature for 1h.

[0131] ② After washing 10 times with 1 mL / L TBS / Tween 20, add 10 μL human phage conformational 7-peptide library (containing 2×10 11 Virus particles), human phage conformation type 7 peptide library including host bacteria ER2738, purchased from NewEngland Biolabs, and adsorbed at room temperature for 50min.

[0132] Wherein the formula of TBS buffer solution is Tris-HCl 50mmol / L, NaCl150mM / L.

[0133] ③ After washing 9 times with 1 mL / L TBS / Tween 20, add 1 mL of 0.2 mol / L HCl-Gly (glycine) elution buffer with a pH value of 2.2 and 1 g / L BSA, dissociate at room temperature for 6 minutes, and then add 150 μL pH 9.1 1mol / L Tris-HCl n...

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Abstract

The present invention discloses a kind of small peptide with the amino acid sequence of P-M-K-M-R-T-M or Y-E-P-K-I-R-G, and its preparation. The preparation process includes screening bacteriophage peptide library with B lymphocyte stimulus factor as target molecule to obtain B lymphocyte stimulus factor specifically combined small peptide and determining its sequence, and subsequent artificial synthesis of corresponding small peptide. The small peptide may be used in preparing medicine for treating B lymphocyte stimulus factor related diseases.

Description

technical field [0001] The invention relates to a small peptide and a screening and preparation method thereof, in particular to a B lymphocyte stimulating factor (BLyS) inhibitory small peptide and a screening and preparation method thereof. Background technique [0002] Systemic lupus erythematosus (SLE), rheumatoid arthritis (RA) and Sjögren's syndrome (SS) are three autoimmune diseases with high prevalence and serious threats to human health (with In my country, for example, the prevalence rate of SLE is 0.07%, and the prevalence rate of women is about 0.1%; the prevalence rate of RA is 0.32%-0.36%; the prevalence rate of SS is 0.3%-0.7%, but the prevalence rate in the elderly population can be as high as 3% to 4%). So far, the treatment measures for the three diseases are mainly the use of anti-inflammatory drugs and immunosuppressants, combined with general treatment of symptomatic treatment. For RA, surgical treatment such as synovectomy or joint replacement can also ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C12P21/02C12Q1/68G01N33/68
Inventor 何凤田吉清郑英如
Owner ARMY MEDICAL UNIV
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