Immunogenic peptides and use thereof

A technology of immunogenicity and immune complexes, applied in the field of diagnosis and pathology, can solve problems such as not being easy to use

Inactive Publication Date: 2018-11-23
OMUNIS +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, although such methods are effective in the early detection of active tuberculosis, they can only be performed in well-equipped l...

Method used

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  • Immunogenic peptides and use thereof
  • Immunogenic peptides and use thereof
  • Immunogenic peptides and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0128] Example 1 - Identification of Peptides to be Screened

[0129] From 25 recombinant proteins of the lipolytic enzyme family, the inventors have performed an epitope screen to rank more than 800 peptides by computer simulation based on different characteristics (such as hydrophobicity, secondary structure, etc.). This sorting allowed the selection of 200 candidates that pooled the most promising multiple features for the diagnosis of active tuberculosis. Each of the 200 peptides was further evaluated for immunogenicity in an ELISA test for screening of these peptides. Based on the results obtained from the patient pool, the 30 best candidates were retained for continued technical evaluation.

[0130] Using computer algorithms and from the provided gene sequences, the main features of the diagnostic candidate proteins were identified and a classification of the most promising peptides was established. In addition to this, the inventors obtained a list of 833 overlapping ...

Embodiment 2

[0151] Example 2 - Reactivity of 4 Peptides on Samples from Different Individual Patients in the Pool Used for Screening

[0152] To confirm that the peptides studied were in fact capable of diagnosing active tuberculosis, the inventors tested 4 peptides (M3:SEQ ID NO:3; C12:SEQ ID NO:2; C2:SEQ ID NO:1 and O7:SEQ ID NO :6) Reactivity.

[0153] The table below shows the results obtained with sera from 16 patients:

[0154] patient

M3

C12

C2

O7

#1

+++

+++

+++

+++

#2

+++

+++

+

+ / -

#3

+++

+ / -

+ / -

+ / -

#4

+++

+++

+ / -

+

#5

+++

+

+

+ / -

#6

+++

+

+ / -

+++

#7

+++

+++

+ / -

+ / -

#8

+++

+ / -

+++

+++

#9

+++

+++

+ / -

-

#10

+++

+++

+++

+++

#11

+

+ / -

+ / -

-

#12

+

+ / -

+ / -

+ / -

#13

+

+

+ / -

-

#14

+ / -

+

+

+

#15

+ / -

-

+ / - ...

Embodiment 3

[0157] Example 3 - Comparison of the reactivity of peptides according to the number of batches of pooled samples used

[0158] In order to assess the reactivity of the peptides and select the most promising peptides for use in the diagnostic test for active tuberculosis, the inventors evaluated the immunogenicity of each peptide on several different pools of samples. Those reactive to all tested sets were selected as the best candidates. The table below shows examples of 3 test peptides in two different pools of positive samples.

[0159] peptide

OD Negative Collection

Positive Set 1 Ratio

Ratio of Positive Set 2

D7

0.303

1.7

1.7

C4

0.077

-0.3

1.8

B4

0.408

0.9

1.0

[0160]At the end of the screen the peptides with a discovery ratio of 1.5 that were the best diagnostic candidates for active TB were selected. Among all peptides tested, some peptides had ratios >1.5 for both pools of patients (Example D7), w...

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Abstract

The invention relates to an in vitro method of screening immunogenic peptides of interest, capable of recognizing antibodies originating from the serum of individuals suffering from active tuberculosis, said method comprising: bringing into contact of peptides of the serum originating from patients suffering from active tuberculosis, detecting the formation of immune complexes in the preceding step, and selecting peptides of interest for which the value of a ratio R is greater than or equal to 1.5, the ratio R being the measurement value of the formation of immune complex to the measurement value obtained from the control sample.

Description

technical field [0001] The present invention relates to immunogenic peptides and their use, especially in the diagnosis of pathologies. Background technique [0002] With a third of the world's population infected and one death every 20 seconds, tuberculosis (TB) remains one of the world's deadliest diseases. There are currently no rapid, accurate diagnostic tests that can be validated against the expectations of health authorities, especially the World Health Organization (WHO), while reference tests take weeks to produce results. [0003] Current international reference methods for diagnosing TB are time-consuming (up to 8 weeks in bacterial culture), inefficient, expensive, and difficult to implement because they use lung specimens, require a safety level 3 laboratory, and require hospitalization. [0004] In 2011, the World Health Organization published for the first time explicit negative general policy recommendations on the use of first-generation serological tests f...

Claims

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Application Information

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IPC IPC(8): G01N33/569C07K14/35
CPCG01N33/5695G01N2333/35C07K14/35G01N2800/26
Inventor P-A·路博艾杜瓦·图艾龙
Owner OMUNIS
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