The Antibody Society's antibody validation webinar series.

In the wake of the reproducibility crisis and numerous discussions on how commercially available antibodies as research tool contribute to it, The Antibody Society developed a series of 10 webinars to address the issues involved. The webinars were delivered by speakers with both academic and commercial backgrounds. This report highlights the problems, and offers solutions to help the scientific community appropriately identify the right antibodies and to validate them for their research and development projects.

The challenges with the validation of research antibodies.

This article further discusses the reproducibility crisis in biomedical science and how poor conduct of commercial antibodies contribute to this. In addition, the way quality data are presented on product sheets by antibody vendors is scrutinized. The article proposes that there is a distinction between testing data and validation data, and special attention is asked for consistency between batches and aliquots.

Genomics, microRNA, epigenetics, and proteomics for future diagnosis, treatment and monitoring response in upper GI cancers.

One major objective for our evolving understanding in the treatment of cancers will be to address how a combination of diagnosis and treatment strategies can be used to integrate patient and tumor variables with an outcome-oriented approach. Such an approach, in a multimodal therapy setting, could identify those patients (1) who should undergo a defined treatment (personalized therapy) (2) in whom modifications of the multimodal therapy due to observed responses might lead to an improvement of the response and/or prognosis (individualized therapy), (3) who might not benefit from a particular toxic treatment regimen, and (4) who could be identified early on and thereby be spared the morbidity associated with such treatments. These strategies could lead in the direction of precision medicine and there is hope of integrating translational molecular data to improve cancer classifications.

Biomarkers and their dependence on well-reported antibodies.

Jan Voskuil is the Chief Scientific Officer at antibody manufacturer Everest Biotech in Oxfordshire, UK. After specializing in prokaryotic cell biology through his PhD program in Amsterdam, The Netherlands and a postdoctorate position at Stanford (CA, USA), he switched to the science of neurodegenerative diseases at Oxford, UK through postdoctorate positions at Dunn School of Pathology and at MRC and through a leading position at the Alzheimer drug discovery company Synaptica. He subsequently gained experience in a Good Laboratory Practice-regulatory environment in contract research organization companies both in Oxfordshire and Cambridgeshire, validating assays in Flow Cytometry and ELISA platforms and writing standard operating procedures.

How difficult is the validation of clinical biomarkers?

Recent developments of introducing stratified medicine/personal health care have led to an increased demand for specific biomarkers. However, despite the myriads of biomarkers claimed to be fit for all sorts of diseases and applications, the scientific integrity of the claims and therefore their credibility is far from satisfactory. Biomarker databases are met with scepticism.

Commercial antibodies and their validation.

Despite an impressive growth in the business of research antibodies a general lack of trust in commercial antibodies remains in place. A variety of issues, each one potentially causing an antibody to fail, underpin the frustrations that scientists endure. Lots of money goes to waste in buying and trying one failing antibody after the other without realizing all the pitfalls that come with the product: Antibodies can get inactivated, both the biological material and the assay itself can potentially be flawed, a single antibody featuring in many different catalogues can be deemed as a set of different products, and a bad choice of antibody type, wrong dilutions, and lack of proper validation can all jeopardize the intended experiments.

Diffuse enteritis after colectomy for ulcerative colitis: two case reports and review of the literature.

Ulcerative colitis (UC) is an inflammatory disease of the colon. Involvement of the small bowel is limited to backwash ileitis or pouch-related conditions. Here, we report two men with UC who presented with small bowel inflammation and even perforation, within 1 month after subtotal colectomy.

DYX1C1 functions in neuronal migration in developing neocortex.

Rodent homologues of two candidate dyslexia susceptibility genes, Kiaa0319 and Dcdc2, have been shown to play roles in neuronal migration in developing cerebral neocortex. This functional role is consistent with the hypothesis that dyslexia susceptibility is increased by interference with normal neural development. In this study we report that in utero RNA interference against the rat homolog of another candidate dyslexia susceptibility gene, DYX1C1, disrupts neuronal migration in developing neocortex.

The intact human acetylcholinesterase C-terminal oligomerization domain is alpha-helical in situ and in isolation, but a shorter fragment forms beta-sheet-rich amyloid fibrils and protofibrillar oligomers.

A 14-residue fragment of the C-terminal oligomerization domain, or T-peptide, of human acetylcholinesterase (AChE) shares sequence homology with the amyloid-beta peptide implicated in Alzheimer's disease and can spontaneously self-assemble into classical amyloid fibrils under physiological conditions [Greenfield, S. A., and Vaux, D.

Occurrence of esophageal granular cell tumors in The Netherlands between 1988 and 1994.

Granular cell tumors (GCT) of the esophagus are rare. The tumor is generally beleived to be of neurogenic origin and shows a malignant course in 2-4% of cases. No unanimity has been reached regarding the management of this tumor.

Biodistribution of 123I-labeled 4-hydroxytamoxifen derivatives in rats with dimethylbenzanthracene-induced mammary carcinomas.

A 123I-labeled tamoxifen derivative with a high affinity for the antiestrogen-binding site (AEBS) has been prepared. Biodistribution studies in rats showed a good linear correlation between the AEBS contents of tissue in fmol/g and the accumulated amount of radioactivity in percent dose per gram at 24 h.

Gastroesophageal reflux disease in intellectually disabled individuals: leads for diagnosis and the effect of omeprazole therapy.

Objectives: The therapeutic approach to gastroesophageal reflux disease (GERD) in intellectually disabled individuals has not been studied extensively. So far, only low response rates to medical and surgical therapy of GERD have been reported. However, the efficacy of proton pump inhibitors, to date the most effective medical therapy for GERD, has never been evaluated in this population.

Prevalence of esophageal disorders in patients with chest pain newly referred to the cardiologist.

Study Objective: The prevalence of esophageal disorders (dysmotility and/or gastroesophageal reflux) in patients with chest pain newly referred to a cardiologic clinic is unknown. The aims of our study were to investigate the prevalence of esophageal abnormalities in these patients and to assess the value of medical history in predicting the origin of the patient's chest pain.

Design: We evaluated 28 consecutive patients who were newly referred to the cardiologist because of angina-like chest pain.

How does FtsZ find its location?

The conformational flexibility of FtsZ and the properties of its epitopes have been studied. Cellular fractions of Escherichia coli have been treated with Triton X-114. FtsZ distributed in the polar as well as in the non-polar phase.

The choriocarcinoma syndrome: an emergency.

A 36-year-old male patient is described who presented with gynaecomastia, pulmonary nodules and a retroperitoneal mass in combination with a markedly elevated HCG level. A diagnosis of "choriocarcinoma syndrome" was made. Despite a clear response from the tumour to chemotherapy the patient died, at least partially due to delay in treatment.

Epitope mapping of Escherichia coli cell division protein FtsZ with monoclonal antibodies.

A fusion between lacZ and ftsZ of Escherichia coli was constructed to obtain a beta-galactosidase-FtsZ fusion protein. This fusion protein was used to raise antibodies against cell division protein FtsZ. Six monoclonal antibodies were obtained, and they reacted with FtsZ from cytoplasm and membrane fractions.

Cefepime compared with ceftazidime as initial therapy for serious bacterial infections and sepsis syndrome.

In an open randomized multicenter comparative study, we evaluated the safety and efficacy of cefepime (CP; 2.0 g given intravenously every 12 h) and ceftazidime (CZ; 2.0 g given intravenously every 8 h) as initial treatment for adult patients with suspected serious bacterial infections.

Calcium in bacteria: a solution to which problem?

Calcium and calcium-binding proteins including those resembling calmodulin are implicated in numerous diverse processes in bacteria. These processes include chemotaxis, sporulation, virulence, the transport of sugars and proteins, phosphorylation, heat shock, the initiation of DNA replication, septation, nucleoid structure, nuclease activity and recombination, the stability of the envelope, and phospholipid synthesis and configuration. That such varied processes should have a common factor, calcium, suggests major underlying principles of calcium metabolism which have yet to be discovered.

Stimulation and inhibition of cell division in synchronized Escherichia coli.

Escherichia coli was synchronized by centrifugal elutriation. When grown in a Tris-based medium, addition of EDTA resulted in division about 20 min earlier (division of control at t = 75 min). EDTA addition caused a change in cell shape, with cells becoming narrower and longer, whereas the surface area to volume ratio increased.