Recombinant Antibodies for Immunotherapy Research

At Absolute Antibody we have developed a collection of recombinant engineered antibodies against clinically relevant proteins, including homologues of current therapeutic targets. We can match the antibody species to the host organism and tailor the effector function, similar to how pharmaceutical companies develop human therapeutics.

t cell receptor

Target (mouse) Clone ID Original Format
CD134 OX86 Rat IgG1
CD200 OX90 Rat IgG2a
CD28 D665 Mouse IgG1
CD28 E18 Mouse IgG2b
CD47 mIAP301 Rat IgG2a
GITR YGITR 860.103.5 Rat IgG2b
GITR YGITR 765 Rat IgG2b
GITRL YGL 386 Rat IgG1
ICOS 7E.17G9 Rat IgG2b
LAG3 YAML 555.6 Rat IgG2b
PD-1 RMP1-14 Rat IgG2a
PD-L1 alphaPD-L1 Chicken scFv
PD-L1 YDC 127.1.1 Rat IgG2a

Antibodies against proteins involved in co-stimulation and other aspects of immune cell regulation are of particular interest to therapeutics developers. Some have already entered the clinic, with more in the development pipeline. However, many aspects of immune cell signalling are still unknown, and researchers require ever more advanced tools to tap into this potential.
At Absolute Antibody, we use recombinant technology to provide superior monoclonal antibody reagents at competitive prices. In particular we can modify antibody species and isotype for greater flexibility in vivo, for example we can readily generate mouse-anti-mouse or rat-anti-rat antibodies.

Why go recombinant?
Because of their recombinant manufacture our antibodies show minimal batch-to-batch variability and have potential for customisation. We can convert any antibody into any format allowing us to offer each specificity in a range of species, isotypes and subtypes. This means our customers may ‘build’ an antibody to best suit their experiment.

  • Choose primary antibody format to suit your secondary reagent
  • Choose antibody species to be compatible with your model organism
  • Choose antibody isotype to investigate your chosen host responses (includes IgM and all IgG subtypes)
  • Choose from a range of custom engineering options such as our Fc Silent format with reduced FcR binding to remove Fc receptor function, or other such formats found in the literature (e.g. IgG1-LALA, IgG1-D265A)
  • Choose one of our listed antibodies, or apply our recombinant technology to your own clone. All our antibody services are royalty-free.

Soluble B7-H1/PD-L1 Levels in Multiple Cancer Subtypes: High Sensitivity Measurement by Immunoassay

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Although the immune checkpoint protein B7-H1/PD-L1 is a commonly used cancer biomarker, controversy remains over the predictive and prognostic utility of immunohistochemical methods. Download our white paper which shows that we can reliably measure B7-H1/PD-L1 in conditioned media supernates and cell lysates from a number of cancer cell lines.

Data from our white paper indicate that:

  • We can reliably measure B7-H1/PD-L1 in a number of human cancer cell lines
  • Soluble B7-H1/PD-L1 expression levels are coincident with loss of PTEN tumor suppressor or increased activation of the phosphoinositide 3-kinase pathway

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Request a copy:

The PD-1 / PD-L1 Pathway

Introduction

The immune system fights off pathogens, but this defensive force can be pathogenic itself when hyperactive, resulting in autoimmune diseases such as lupus and multiple sclerosis. Consequently, the body has developed multiple mechanisms to suppress the immune system when necessary.

One method of immunosuppression is the PD-1 pathway. This pathway is activated in response to the mobilization of the immune system. The receptor PD-1 is expressed on the surface of activated lymphocytes. Similarly, its ligand, PD-L1, is expressed by antigen-presenting cells in response to cytokine signaling. When PD-L1 is bound to PD-1, downstream signaling undoes the phosphorylation events associated with activation, thereby reverting lymphocytes to an inactive state [1, 2].

Antibody Cat no. Type Applications
PD-1/CD279 Antibody 18106-1-AP Rabbit Poly ELISA, WB, FC, IHC
pdd1
IHC of paraffin-embedded human ovary tumor tissue slide using PD-1/CD279 Antibody,at dilution of 1:200 (under 10x lens).

Immunotherapy: A Promising Treatment for Cancer

Tumor cells take advantage of the PD-1 pathway to evade the immune system [3]. Consequently, many pharmaceutical companies have been developing drugs to inhibit PD-1 and PD-L1. In clinical trials, many patients have shown strong responses to these therapies [4-10]. For these drugs to be most effective, a high number of CD8+ T cells must already be at the tumor site, ready to be mobilized after inhibition of the PD-1 pathway [11].Recent investigations have uncovered promising methods of increasing the efficacy of PD1 inhibitors. One method is combining PD1 inhibitors with other drugs, such as HDAC inhibitors [12] and anti-CTLA4 antibodies [9]. Another method is using biomarkers to predict response to therapy [13,14]. Recent work has also suggested that GSK3 inhibitors can enhance effects of immunotherapy [15].Though these results have been encouraging, several challenges remain, including the mitigation of autoimmune effects and how to overcome drug resistance.

Current Antibody Drug Development

Drug Company
PD-1 SHR-1210 Incyte
Nivolumab Bristol-Myers Squibb
Pembrolizumab Merck
Pidilizumab CureTech
BMS 936559 Bristol-Myers Squibb
PD-L1 Atezolizumab Roche
Durvalumab AstraZeneca
Avelumab Pfizer/Merck
MDX-1105 Bristol-Myers Squibb
CTLA-4 Ipilimumab Bristol-Myers Squibb
Tremelimumab Pfizer/AstraZeneca
Antibody Cat no. Type Applications
PD-L1/CD274 Antibody 17952-1-AP Rabbit Poly ELISA, IF, IHC, IP, WB

KD/KO Validated, 9 Publications

pdl1
WB result of PD-L1 antibody (17952-1-AP, 1:500) with si-Control and si-PD-L1 transfected HepG2 and HeLa cells with 3 separate constructs.

Related Products

Antibody Cat no. Type Applications
PD-L1/CD274  66248-1-Ig  Mouse mono  ELISA, WB, IHC, IF, FC
CD86/CTLA 4  13395-1-AP  Rabbit poly  ELISA, WB, FC
CD3 epsilon  17617-1-AP  Rabbit poly  ELISA, WB, IHC, IP, FC
GSK3B  22104-1-AP  Rabbit poly  ELISA, WB, IHC, IF, IP
BRAF  20899-1-AP  Rabbit poly  ELISA, IHC, IF

References

1. Ohegbulam et al. (2015) Human cancer immunotherapy with antibodies to the PD-1 and PD-L1 pathway. Trends Mol Med. 21:24-33.

2. Haanen, J. (2013) Immunotherapy of Melanoma. EJC Suppl 11:97-105.

3. Yao et al. (2013) Advances in targeting cell surface signaling molecules for immune modulation. Nat Rev Drug Discov. 12:130-146.

4. Topalian, S. et al. (2012) Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. N. Engl. J. Med. 366:2443-2454.

5. Hamid, O. et al. (2013) Safety and tumor responses with lambrolizumab (anti-PD-1) in melanoma. N. Engl. J. Med. 369:134-144.

6. Topalian, S. L. et al. (2012) Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. N. Engl. J. Med. 366:2443–2454

7. Brahmer, J. R. et al. (2012) Safety and activity of anti-PD-L1 antibody in patients with advanced cancer. N. Engl. J. Med. 366:2455–2465

8. Hamid, O. et al. (2013) Safety and tumor responses with lambrolizumab (anti-PD-1) in melanoma. N. Engl. J. Med. 369:134–144

9. Wolchok, J. D. et al. (2013) Nivolumab plus ipilimumab in advanced melanoma. N. Engl. J. Med. 369:122–133

10. Topalian, S. L. et al. (2014) Survival, durable tumor remission, and long-term safety in patients with advanced melanoma receiving nivolumab. J. Clin. Oncol. 32:1020–1030

11. Tumeh, P. et al. (2014) PD-1 blockade induces responses by inhibiting adaptive immune response. Nature. 515:568-71.

12. Woods, D. et al. (2015) HDAC inhibition upregulates PD-1 ligands in melanoma and augments immunotherapy with PD-1 blockade. Cancer Immunol Res 12:1375-85.

13. Chakravarti, N., & Prieto, V. G. (2015). Predictive factors of activity of anti-programmed death-1/programmed death ligand-1 drugs: immunohistochemistry analysis. Translational Lung Cancer Research, 4:743–751.

14. Barak, V. et al. (2015) Assessing response to new treatments and prognosis in melanoma patients, by the biomarker S-100B. Anticancer Res. 35:6755-60.

15. Taylor, A. et al. (2014) Glycogen synthase kinase 3 inactivation drives T-bet-mediated downregulation of co-receptor PD-1 to enhance CD8+ cytolytic T cell responses. Immunity 44:274-86.

Rapid isolation of untouched neurons

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Using our Neuron Isolation Kit, you can now rapidly isolate viable and pure neurons from both neonatal and adult mouse brain by depletion of non-neuronal cells. The high purity of the isolated neurons relies on the specificity of the non-neuronal cell surface marker used in the Neuron Isolation kit.

Download this free scientific poster today and see scientific data about the expression of the non-neuronal cell marker:

  • In different neural cell types
  • At different mouse brain regions
  • During different developmental stages (from E14 to P7)

DOWNLOAD THE FREE POSTER

New mGlu Receptor Agonists and Antagonists

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Tocris’s new selection of mGlu receptor ligands.

Key Features of (±)-ADX 71743

  • Negative allosteric modulator at mGlu7
  • Blocks high frequency-induced LTP at SC-CA1 synapses in hippocampal slices
  • Increases evoked excitatory postsynaptic currents (eEPSCs) in mice
  • Brain penetrant

Please click on the product names below to view activity data, references and purity information.

Cat. No. Product Name Description
5377 VU 0483605 Positive allosteric modulator at mGlu1
5613 AZD 9272 Potent and selective mGlu5 antagonist
5614 AZD 2066 mGlu5 antagonist; brain penetrant
5693 VU 0409551 Selective mGlu5 positive allosteric modulator
5715 (±)-ADX 71743 Potent and selective NAM at mGlu7 receptors

View our full range of mGlu receptor ligands.

Monoclonal antibody with conformational specificity for a toxic conformer of amyloid β42

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Amyloid β-protein (Aβ42) oligomerization is an early event in Alzheimer’s disease (AD). Current diagnostic methods using sequence-specific antibodies against less toxic fibrillar and monomeric Aβ42 run the risk of overdiagnosis. Hence, conformation-specific antibodies against neurotoxic Aβ42 oligomers have garnered much attention for developing more accurate diagnostics. Antibody 24B3, highly specific for the toxic Aβ42 conformer that has a turn at Glu22 and Asp23, recognizes a putative Aβ42 dimer, which forms stable and neurotoxic oligomers more potently than the monomer. 24B3 significantly rescues Aβ42-induced neurotoxicity, whereas sequence-specific antibodies such as 4G8 and 82E1, which recognizes the N-terminus, do not. The ratio of toxic to total Aβ42 in the cerebrospinal fluid of AD patients is significantly higher than in control subjects as measured by sandwich ELISA using antibodies 24B3 and 82E1. Thus, 24B3 may be useful for AD diagnosis and therapy.

Monoclonal antibody with conformational specificity for a toxic conformer of amyloid β42 and its application toward the Alzheimer’s disease diagnosis. Murakami K et al. Sci Rep. 2016 Jul 4;6:29038 PMID: 27374357

#27709 Human Amyloidβ Toxic Oligomer Assay Kit – IBL is developed using the antibody (clone: 24B3) specifically detects a toxic Amyloid Beta conformer. It measures selectively putative Amyloid Beta Oligomer in CSF.

Therapeutic ANGPTL2 suppression could antagonize development of heart failure

The research group led by Professor Yuichi Oike at Kumamoto University reported that expression of angiopoietin-like protein 2 (ANGPTL2) increases in pathologically-remodeled hearts of mice and humans, while decreased cardiac ANGPTL2 expression occurs in physiological cardiac remodelling induced by endurance training in mice. Mice overexpressing ANGPTL2 in heart show cardiac dysfunction caused by both inactivation of AKT and sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA)2a signalling and decreased myocardial energy metabolism. Conversely, Angptl2 knockout mice exhibit increased left ventricular contractility and upregulated AKT-SERCA2a signalling and energy metabolism. Finally, ANGPTL2-knockdown in mice subjected to pressure overload ameliorates cardiac dysfunction. Overall, these studies suggest that therapeutic ANGPTL2 suppression could antagonize development of heart failure.

ANGPTL2 activity in cardiac pathologies accelerates heart failure by perturbing cardiac function and energy metabolism. Tian Z et al. Nat Commun. 2016 Sep 28;7:13016.
PMID: 27677409

#27745 Human ANGPTL2 Assay Kit – IBL was used in this study.