Products

PepMix™


The use of complete protein-spanning mixtures of overlapping peptides is extremely efficient for immunostimulation of T-lymphocytes and diagnostic applications. JPT Peptide Technologies offers cost-effective ready-to-use PepMixes™ on a per-test basis. In addition, customized PepMixes™ will be prepared in a fast and economical manner according to the specific needs of the customer.

In order to establish reliable and validated T-cell assays such as ELISPOT, appropriate positive and negative controls are essential. Selection of such controls helps to confirm proper functionality of the assay as well as viability of the cells used. In addition, high quality controls are most important to establish standardized assay protocols in clinical setups. Compared to commonly used controls like phytohemagglutinin (PHA) and concanavalin A (ConA) that stimulate T-cells as well as other cell types or full length antigens, synthetic peptide pools offer the advantage of a high batch-to-batch reproducibility, applicability of reliable chemical and biochemical QC/QA measures and shelf stability when stored as freeze dried aliquots.

JPT offers several synthetic peptide pools that can serve as positive controls by stimulating antigen specific CD8+ T cell responses (CEF pools) or both CD4+ & CD8+ T-cell responses (HCMV pp65, IE-1, IE-2 pools). Whereas our CEF pools contain 23 (standard) or 32 (extended) different class-I restricted peptides deducted from CMC, EBV and Influenca virus, our CMV peptide pools are constructed from 15meric peptide scans overlapping by 11 amino acids through full length antigens. Thus, the latter pools are able to stimulate both CD4+ and CD8+ responses within a single control experiment.

In addition, JPT offers a variety of peptide pool based negative controls that offer a complement to the commonly used protocols that make use of cells in medium without respective antigen stimulation.


PepMixes™ of the following proteins are available in stock:

Positive control pools for T-Cell Assays


PepMix Product Code Protein Source Primary accession
number
Peptides 
HCMVA (IE-1) PM-IE1 55 kDa immediate-early protein 1 (VIE1_HCMVA) Human cytomegalovirus P13202 120
HCMVA (IE-2) PM-IE2 45 kDa immediate-early protein 2 (VIE2_HCMVA) Human cytomegalovirus P19893 143
HCMVA (pp65) PM-PP65 65 kDa lower matrix phosphoprotein (PP65_HCMVA) Human cytomegalovirus P06725 138
CEF Pool (standard)    PM-CEF-S defined HLA class I-restricted
T-cell epitopes
Human cytomegalovirus,
Epstein-Barr virus, Influenza virus
n.a. 23 
CEF Pool (extended)       PM-CEF-E defined HLA class I-restricted
T-cell epitopes
Human cytomegalovirus,
Epstein-Barr virus, Influenza virus
n.a. 32

Negative control pools for T-Cell Assays


PepMix Product Code Protein Source Primary accession
number
Peptides 
ACTS_Human PM-ACTS Alpha-actin-1 Homo Sapiens P68133 92
HLA classI
Ig-like C1 domain
PM-C1 Human HLA n.a. 19
HIV (Con B gag motif) PM-HIVgag Con B gag motif Human immunodefficiency virus n.a. 123
MOG Human
(aa 30 -154)
PM-MOG Myelin-Oligo
dentrocyte glycoprotein
Homo Sapiens Q16653 29

Infectious Diseases


PepMix Product Code Protein Source Primary accession
number
Peptides 
BKV
(capsid protein VP1)
PM-BKV-VP1 Capsid protein VP1 (VP1_POVBA) BK polyomavirus P14996 88
BKV
(capsid protein VP2)
PM-BKV-VP1 Capsid protein VP2 (VP2_POVBK) BK polyomavirus P03094 85
BKV (VP3) PM-BKV-VP3 Capsid protein VP2 (VP3 isoform) BK polyomavirus P03094-2 56
BKV
(large T antigen)
PM-BKV-LTA Large T antigen (LT1_POVBA) BK polyomavirus P14999 170
BKV
(small T antigen)
PM-BKV-StA Small T antigen (ST1_POVBK) BK polyomavirus P03082 41
Candida (MP65) PM-CAND-MP65 Mannoprotein MP Candida albicans Q9HEP1 92
EBV (BZLF1) PM-EBV-BZLF1 Trans-activator protein BZLF1 (BZLF1_EBV) Epstein-Barr virus P03206 59
EBV (EBNA1) PM-EBV-EBNA1 Epstein-Barr virus Nuclear Antigen Epstein-Barr virus YP401677 158
EBV (EBNA3a) PM-EBV-EBNA3a Epstein-Barr virus Nuclear Antigen Epstein-Barr virus YP_401669  234
EBV (EBNA3b) PM-EBV-EBNA3b Epstein-Barr virus Nuclear Antigen Epstein-Barr virus ABB89244 279
EBV (EBNA3c) PM-EBV-EBNA3c Epstein-Barr virus Nuclear Antigen Epstein-Barr virus ABB89245 265
EBV (LMP1) PM-EBV-LMP1 Latent membrane protein 1 (LMP1_EBV) Epstein-Barr virus P03230 94
EBV (LMP2) PM-EBV-LMP2 Latent membrane protein 2 (LMP2_EBV) Epstein-Barr virus P13285 122
HAdV-3
(hexon protein)
PM-HadV3 Hexon protein (HEX_ADE03) Human adenovirus 3 P36849 234
HAdV-5
(penton protein)
PM-HadV5 Penton protein (PEN3_ADE05) Human adenovirus 5 P12538 140
HBV
(large env.protein)
PM-HBV-lEV Large surface protein Hepatitis B virus P17101 98
HCMVA (IE-1) PM-IE1 55 kDa immediate-early protein 1 (VIE1_HCMVA) Human cytomegalovirus P13202 120
HCMVA (IE-2) PM-IE2 45 kDa immediate-early protein 2 (VIE2_HCMVA) Human cytomegalovirus P19893 143
HCMVA (pp65) PM-pp65 65 kDa lower matrix phosphoprotein (PP65_HCMVA) Human cytomegalovirus P06725 138
HCMVA (UL40) PM-UL40 Uncharacterized protein UL40 (UL40_HCMVA) Human cytomegalovirus P16780 53
HHV6 U54 PM-HHV-U54 U54 Human Herpesvirus 6B Q9QJ29 112
HIV-1
(Con B gag motif)
PM-HIVgag Con B gag motif Human immunodeficiency
virus
n.a. 123
Influenza A
(MP1 protein)
PM-INFA-MP1 Membrane protein M1 Influenza A virus AAA43277 110
Influenza A
(NP protein)
PM-INFA-NP Nucleocapsid protein Influenza A virus ABB79814 61
RSV (nucleocapsid protein N)  PM-RSV-NCPN Nucleocapsid protein Human respiratory syncytial virus  AAB82431 95
RSV (protein F) PM-RSV-FGF0 Fusion glycoprotein F0 Human respiratory syncytial virus  036634  141
Yellow fever NS4B PM-YF-NS4B Genome polyprotein Yellow fever virus PO3314 60

Cancer


PepMix Product Code Protein Source Primary accession
number
Peptides 
CEA PM-CEA Carcinoembryonic antigen Homo sapiens P06731   173
Claudin-6 PM-Cl-6 Claudin-6 Homo Sapiens P56747 53
c-Myc (MYC_Human) PM-cMyc Myc proto-oncogene Homo sapiens P01106 107
Cyclin B1 PM-CYC-B1 CCNB1 Homo sapiens   P14635 106
ERBB2_ECD PM-ERB_ECD ERBB2 (-1 1-683) Homo sapiens P04626 168
HER 2 ICD PM-ERB-ICD ERBB2 (-1 662-1255) Homo sapiens  P04626  146
Histone H1.2 PM-Histone H1 HIST1H1C Homo sapiens P16403 51
Histone H4_Human PM-H4 Histone H4 Homo sapiens  P62805  23
Melan-A/MART-1      PM-MelA MELAN-A protein (MAR1_HUMAN)           Homo sapiens                Q16655 27
MAGEA1 PM-MAGEA1 Melanoma-associated antigen 1 Homo sapiens P43355 75
MAGEA3 PM-MAGEA3 Melanoma-associated antigen 3 Homo sapiens P43357  76
MAGEA4 PM-MAGEA4 Melanoma antigen family A, 4 Homo sapiens P43358 77
Mammaglobin A PM-MamA Mammaglobin A (SG2A2_human) Homo sapiens Q13296 21
MMP11 PM-MMP11 Stromelysin 3 Homo sapiens P24347 121
MVA 018L PM-MVA_018L Host range protein 2 Vaccinia virus P68598 35
MVA 074R PM-MVA-074R Putative 49.8k protein Vaccinia virus  O57196  106
MVA 093L PM-MVA-093L IMV associated protein p35 Vaccinia virus O57206  79
MVA 105L PM-MVA-105L Cell-surface binding protein Vaccinia virus O57211 74
MVA 121L (1+2) PM-MVA-121L Major Core protein p4a Vaccinia virus O57223  111+112
MVA 189R PM-MVA-189R Putative 21.7k protein Vaccinia virus O57265 45
NY-ESO-1 PM-NYE Cancer/testis antigen 1 (CTG1B_HUMAN)   Homo sapiens P78358 43
P53_human PM-p53 Cellular Tumor antigen p53 Homo sapiens  P04637  96
PRAME/OIP4 PM-OIP4 Melanoma antigen preferentially
expressed in tumors (MAPE_HUMAN)
Homo sapiens P78395 125
PSA (human) PM-PSA Prostate-specific antigen Homo sapiens P07288 63
SOX-2 PM-SOX-2 Transcription factor SOX-2 Homo sapiens P48431 77
SSX2 PM-SSX2 SSX2 Homo sapiens Q16385 45
Survivin_BIRC5 PM-Survivin Baculoviral IAP repeat-containing
protein 5
Pongo abelii
(Sumatran orangutan)
Q5RAH9  33
TARP PM-TARP TCR gamma alternate reading frame
protein isoform 1
Homo sapiens NP_001003799 12
TRP 2 PM-TRP-2 L-dopachrome tautomerase Homo sapiens P40126 127
Tyrosinase_human PM-Tyr_H Tyrosinase Homo sapiens P14679  117
Tyrosinase_mouse PM-Tyr_M Tyrosinase Mus musculus (mouse) P11344 117
WRKY 47 PM-WRK Probable WKRY transcription factor 47 Arabidopsis thaliana  Q9ZS17  120
WT1 (WT33) PM-WT1 Wilms tumor protein (WT1_HUMAN) Homo sapiens P19544 110

Custom PepMixes™

Custom PepMixes are available via our High Throughput Custom Synthesis program. Protein primary sequences provided will be chemically synthesized as overlapping peptides, purified and analyzed to comply with the requirements of T-cell assays. All production steps are optimized to avoid contaminations and failure sequences which may lead to false positive T-cell responses or inhibition of T-cell responses.

Pooling and aliquotation will be performed using validated protocols ensuring the presence of all peptides in the respective pools. Our fully automated aliquotation service yields freeze dried pool aliquots avoiding deterioration of peptides in DMSO stock solutions.

Custom PepMix Production


Applications for PepMix™

  • Immunostimulation of T-lymphocytes
  • Monitoring of immune status during diseases
  • Assessment of vaccine efficacy

Benefits of PepMix™

  • Equivalent or better stimulation of CD4 and CD8
  • T-cell responses compared to whole protein antigens
  • Simultaneous detection of CD4 and CD8 responses in a single sample
  • Improved responses in stored blood and PBM cells compared to whole protein antigens

Single Peptides

In addition to the mixes, peptides can also be ordered as single peptide aliquots to validate the results found using the mixes. All peptides and mixes will be quality controlled to ensure batch-to-batch reproducibility!

Testimonials

"I have used JPT's peptide pools (PepMix) for years now, with great satisfaction! First CMV derived peptides and now also EBV. So far we have published data in one publication and plan several more in the near future."

Anna Karin Lidehaell (Uppsala University, Clin. Immunol., Uppsala, Sweden)



"The focus of the Clinical Immunology Group at the German Rheumatism Research Centre is to study the role of cells and molecules in the origin and progression of autoimmune and other diseases. In our hands PepMixes™ were found to be effective in monitoring the immune status of various patient populations but show also promise for the development of novel immuno therapy approaches."

Andreas Thiel, PhD (German Rheumatism Research Centre, Clin. Immunol., Berlin, Germany)

 


Selected References:

  • Clonotype Analysis of Cytomegalovirus-Specific Cytotoxic T Lymphocytes 
    Babel et al., Journal of the American Society of Nephrology (2008) (abstract)
  • Prophylactic Transfer of CD8-Depleted Donor Lymphocytes after T-cell Depleted Reduced-Intensity Transplantation
    Meyer et al., Blood (2008) (abstract)
  • Massive Load of Functional Effector CD4+ and CD8+ T Cells against Cytomegalovirus in Very Old Subjects
    Myers et al., J. Immunol. (2007) (abstract)
  • Reconstitution of Adenovirus-Specific Cell-mediated immunity in pediatric patients after Hematopoietic Stem Cell Transplantation
    Myers et al., Bone Marrow Transplant. (2007) (abstract)
  • Monoculture-Derived T Lymphocytes Specific for Multiple Viruses Expand and Produce Clinically Relevant Effects in Immunocompromised Individuals
    Leen at al., Nat. Med. (2006) (abstract)
  • Results of a Cytomegalovirus (CMV)-Specific CD8+/Interferon- Gamma+ Cytokine Flow Cytometry Assay Correlate with Clinical Evidence of Protective Immunity in Patients with AIDS with CMV Retinitis
    Jacobson et al., J. Infect. Dis. (2004) (abstract)
  • Functional Comparison of T Cells Recognizing Cytomegalovirus pp65 and Intermediate-Early Antigen Polypeptides in Hematopoietic Stem-Cell Transplant and Solid Organ Transplant Recipients
    Lacey et al., J. Infect. Dis. (2006) (abstract)
  • Acquisition of Direct Antiviral Effector Functions by CMV-Specific CD4+ T Lymphocytes with Cellular Maturation
    Casazza et al., J. Exp. Med. (2006) (abstract)
  • Protection from Cytomegalovirus After Transplantation is Correlated with Immediate Early 1–specific CD8 T cells
    Bunde et al., Eur. J. Exp. Med. (2005) (abstract)
  • HLA Type-Independent Generation of Antigen-Specific T Cells for Adoptive Immunotherapy
    Hammer et al., Eur. J. Immunol. (2005) (abstract)
  • Generation of Cytomegalovirus (CMV)-Specific T-lymphocytes Using Protein-Spanning Pools of pp65-Derived Overlapping Pentadecapeptides for Adoptive Immunotherapy
    Trivedi et al., Blood (2005) (abstract)
  • Confirmation of Mycobacterium Tubercolosis Infection by Flow Cytometry After Ex Vivo Incubation of Peripheral Blood T Cells with an ESAT-6-Derived Peptide Pool
    Tesfa et al., Cytometry Part B (Clinical Cytometry) (2004) (abstract)
  • CMV Antigen-Specific CD4+ and CD8+ T Cell IFN Expression and Proliferation Responses in Healthy CMV-Seropositive Individuals
    Sinclair et al., Viral Immunol. (2004), (abstract)
  • Recent Advances in the Development of HIV-1 Vaccines Using Replication-Incompetent Adenovirus Vectors
    Shiver and Emini, Annual Review of Medicine (2004), (abstract)
  • Mapping T Cell Epitopes by Flow Cytometry
    Hoffmeister et al., Methods (2003), (abstract)
  • Cytomegalovirus (CMV) Phosphoprotein 65 Makes a Large Contribution to Shaping the T Cell Repertoire in CMV-Exposed Individuals
    Kern et al., J. Infect. Dis.(2002), (abstract)
  • Use of Overlapping Peptide Mixtures as Antigens for Cytokine Flow Cytometry
    Maecker et al., J. Immunol. Methods (2001), (abstract)
  • Analysis of CD8 T Cell Reactivity to Cytomegalovirus Using Protein-Spanning Pools of Overlapping Pentadecapeptides
    Kern et al., Eur. J. Immunol. (2000), (abstract)
  • Putative Immunodominant Human Immunodeficiency Virus-specific CD8+ T-Cell Responses Cannot Be Predicted by Major Histocompatibility Complex Class I Haplotype
    Betts et al., J. Virol. (2000), (abstract)
  • Identification of T-cell Epitopes Using ELISpot and Peptide Pool Arrays, Tobery and Caulfield
    Methods in Molecular Medicine : Molecular Diagnosis of Infectious Deseases, 2/e; Edited by J.Decker and U. Reischl; Humana Press Inc, Totowa, NJ, (abstract)

    More references under JPT Publications/Literature

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