Ribosome Display Technology: Applications in Disease Diagnosis and Control

Antibody ribosome display remains one of the most successful in vitro selection technologies for antibodies fifteen years after it was developed. The unique possibility of direct generation of whole proteins, particularly single-chain antibody fragments (scFvs), has facilitated the establishment of this technology as one of the foremost antibody production methods. Ribosome display has become a vital tool for efficient and low-cost production of antibodies for diagnostics due to its advantageous ability to screen large libraries and generate binders of high affinity. The remarkable flexibility of this method enables its applicability to various platforms. This review focuses on the applications of ribosome display technology in biomedical and agricultural fields in the generation of recombinant scFvs for disease diagnostics and control.

Download Full-text

A New Approach for Rapidly Reshaping Single-Chain Antibody In Vitro by Combining DNA Shuffling with Ribosome Display

The Journal of Biochemistry ◽

10.1093/jb/mvh083 ◽

2004 ◽

Vol 136 (1) ◽

pp. 19-28 ◽

Cited By ~ 11

Author(s):

X.-B. Wang

Keyword(s):

Dna Shuffling ◽

Single Chain ◽

Single Chain Antibody ◽

Ribosome Display ◽

New Approach

Download Full-text

Single-chain Antibody Against Reg4 Suppresses Gastric Cancer Cell Growth and Enhances 5-FU-induced Cell Death in vitro

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520619666181122104720 ◽

2019 ◽

Vol 19 (5) ◽

pp. 610-619 ◽

Cited By ~ 3

Author(s):

Xue-Qing Zhang ◽

Lu-Ting Yu ◽

Pei Du ◽

Tian-Qi Yin ◽

Zhi-Yuan Zhang ◽

...

Keyword(s):

Gastric Cancer ◽

Cell Proliferation ◽

Cell Death ◽

Cancer Cell ◽

Gastric Cancer Cell ◽

Single Chain ◽

Single Chain Antibody ◽

Ags Cells ◽

Thiazolyl Blue Tetrazolium Bromide

Background:Regenerating islet-derived gene family member 4 (Reg4), a well-investigated growth factor in the regenerative pancreas, has recently been reported to be highly associated with a majority of gastrointestinal cancers. Pathological hyper-expression or artificial over-expression of Reg4 causes acceleration of tumor growth, migration, and resistance to chemotherapeutic 5-Fluorouracil (5-FU). Until now, no method has been successfully established for eliminating the effects of Reg4 protein.Methods:This study reports the production of an engineered immunoglobin, a single-chain variable fragment (scFv-Reg4), to specifically bind Reg4 and block the bioactivity. The complementary-determining regions (CDRs) against Reg4 were assigned using MOE and ZDOCK servers. The binding affinity (KD) was determined by bio-layer interferometry (BLI). MKN45 and AGS cell proliferation was determined by Thiazolyl blue tetrazolium bromide (MTT) method and the cell apoptosis was detected by flow cytometry assay.Results:The KD of scFv-Reg4 to Reg4 was determined to be 1.91×10-8. In MKN45 and AGS cell lines, scFv- Reg4 depressed Reg4-stimulated cell proliferation and the inhibitory rates were 27.7±1.5% and 17.3±2.6%, respectively. Furthermore, scFv significantly enhanced 5-FU-induced cell death, from 23.0±1.0% to 28.4±1.2% in MKN45 and 28.2±0.7% to 36.6±0.6% in AGS cells. Treatment with scFv alone could lyse cancer cells to a certain extent, but no significance has been observed.Conclusion:The single-chain antibody (scFv-Reg4) significantly inhibited gastric cancer cell proliferation and synergistically enhanced the lethal effect of 5-FU. Thus, traditional chemo-/radio- therapeutics supplemented with scFv-Reg4 may provide advances in the strategy for gastrointestinal cancer treatment.

Download Full-text

Designing of an Advanced Compression Bioreactor with an Implementation of a Low-Cost Controlling System Connected to a Mobile Application

Processes ◽

10.3390/pr9060915 ◽

2021 ◽

Vol 9 (6) ◽

pp. 915

Author(s):

Gözde Dursun ◽

Muhammad Umer ◽

Bernd Markert ◽

Marcus Stoffel

Keyword(s):

Mobile Application ◽

Low Cost ◽

Experimental Information ◽

Raspberry Pi ◽

Tissue Constructs ◽

Cost Controlling ◽

Controlling System ◽

And Control ◽

Tissue Models

(1) Background: Bioreactors mimic the natural environment of cells and tissues by providing a controlled micro-environment. However, their design is often expensive and complex. Herein, we have introduced the development of a low-cost compression bioreactor which enables the application of different mechanical stimulation regimes to in vitro tissue models and provides the information of applied stress and strain in real-time. (2) Methods: The compression bioreactor is designed using a mini-computer called Raspberry Pi, which is programmed to apply compressive deformation at various strains and frequencies, as well as to measure the force applied to the tissue constructs. Besides this, we have developed a mobile application connected to the bioreactor software to monitor, command, and control experiments via mobile devices. (3) Results: Cell viability results indicate that the newly designed compression bioreactor supports cell cultivation in a sterile environment without any contamination. The developed bioreactor software plots the experimental data of dynamic mechanical loading in a long-term manner, as well as stores them for further data processing. Following in vitro uniaxial compression conditioning of 3D in vitro cartilage models, chondrocyte cell migration was altered positively compared to static cultures. (4) Conclusion: The developed compression bioreactor can support the in vitro tissue model cultivation and monitor the experimental information with a low-cost controlling system and via mobile application. The highly customizable mold inside the cultivation chamber is a significant approach to solve the limited customization capability of the traditional bioreactors. Most importantly, the compression bioreactor prevents operator- and system-dependent variability between experiments by enabling a dynamic culture in a large volume for multiple numbers of in vitro tissue constructs.

Download Full-text

Proanthocyanidins against Oxidative Stress: From Molecular Mechanisms to Clinical Applications

BioMed Research International ◽

10.1155/2018/8584136 ◽

2018 ◽

Vol 2018 ◽

pp. 1-11 ◽

Cited By ~ 12

Author(s):

Lingyu Yang ◽

Dehai Xian ◽

Xia Xiong ◽

Rui Lai ◽

Jing Song ◽

...

Keyword(s):

Oxidative Stress ◽

Signaling Pathways ◽

Molecular Mechanisms ◽

Metabolic Diseases ◽

Low Cost ◽

Natural Agents ◽

Molecular Damage ◽

And Control

Proanthocyanidins (PCs) are naturally occurring polyphenolic compounds abundant in many vegetables, plant skins (rind/bark), seeds, flowers, fruits, and nuts. Numerousin vitroandin vivostudies have demonstrated myriad effects potentially beneficial to human health, such as antioxidation, anti-inflammation, immunomodulation, DNA repair, and antitumor activity. Accumulation of prooxidants such as reactive oxygen species (ROS) exceeding cellular antioxidant capacity results in oxidative stress (OS), which can damage macromolecules (DNA, lipids, and proteins), organelles (membranes and mitochondria), and whole tissues. OS is implicated in the pathogenesis and exacerbation of many cardiovascular, neurodegenerative, dermatological, and metabolic diseases, both through direct molecular damage and secondary activation of stress-associated signaling pathways. PCs are promising natural agents to safely prevent acute damage and control chronic diseases at relatively low cost. In this review, we summarize the molecules and signaling pathways involved in OS and the corresponding therapeutic mechanisms of PCs.

Download Full-text

Use of Low Cost, Low Fidelity Mockups for Preproduction Testing

Proceedings of the Human Factors Society Annual Meeting ◽

10.1177/154193128302700715 ◽

1983 ◽

Vol 27 (7) ◽

pp. 589-591

Author(s):

Carole A. Bohn

Keyword(s):

New Technologies ◽

Low Cost ◽

Rapid Development ◽

Test Methods ◽

Current Approach ◽

Operating Modes ◽

Test And Evaluation ◽

Methods Development ◽

Display Technology ◽

And Control

The new technologies proposed and/or retrofitted into Navy crewstations have demonstrated increasing sophistication and flexibility. Additionally, the crewstation technologies have shown very rapid development cycles. The current approach of reliance solely on flight testing has proven inadequate because of the multitude of equipment operating modes, lack of experimental control of situational variables, possible location/placement of components, variety of operational environments, dynamic crew tasking, and control/display technology unique characteristics. Test methods and relevent criteria are lacking. A quick fix is the use of low fidelity mockups for rapid testing and methods development. Such an approach can be both effective with respect to test dollars and responsive to the dynamics of the control/display development cycle. The present paper discusses the use of the low fidelity simulation in two specific developments. The first example presents the design of formats for a universal control/display layout to be used as a replacement for conventional pushbutton technology. The second example presents testing designed to determine the amount and type of control/display required for a crewstation functional upgrade. Both examples are from the test and evaluation work being performed on Navy patrol aircraft. Finally, a laboratory will be described which is being developed to permit this approach to testing.

Download Full-text

In vitro phage display in a rat beta cell line: a simple approach for the generation of a single-chain antibody targeting a novel beta cell-specific epitope

Diabetologia ◽

10.1007/s00125-010-1725-9 ◽

2010 ◽

Vol 53 (7) ◽

pp. 1384-1394 ◽

Cited By ~ 11

Author(s):

S. Ueberberg ◽

D. Ziegler ◽

W. Schechinger ◽

J. W. Dietrich ◽

S. Akinturk ◽

...

Keyword(s):

Phage Display ◽

Beta Cell ◽

Cell Line ◽

Simple Approach ◽

Single Chain ◽

Single Chain Antibody ◽

Specific Epitope ◽

Beta Cell Line ◽

Antibody Targeting

Download Full-text

Llama-Derived Single-Chain Antibody Fragments Directed to Rotavirus VP6 Protein Possess Broad Neutralizing Activity In Vitro and Confer Protection against Diarrhea in Mice

Journal of Virology ◽

10.1128/jvi.00436-08 ◽

2008 ◽

Vol 82 (19) ◽

pp. 9753-9764 ◽

Cited By ~ 74

Author(s):

Lorena Garaicoechea ◽

Aurelien Olichon ◽

Gisela Marcoppido ◽

Andrés Wigdorovitz ◽

Marina Mozgovoj ◽

...

Keyword(s):

Capsid Protein ◽

Binding Capacity ◽

Antibody Fragments ◽

Target Antigen ◽

Single Chain ◽

Single Chain Antibody ◽

Group A Rotavirus ◽

Group A

ABSTRACT Group A rotavirus is one of the most common causes of severe diarrhea in human infants and newborn animals. Rotavirus virions are triple-layered particles. The outer capsid proteins VP4 and VP7 are highly variable and represent the major neutralizing antigens. The inner capsid protein VP6 is conserved among group A rotaviruses, is highly immunogenic, and is the target antigen of most immunodiagnosis tests. Llama-derived single-chain antibody fragments (VHH) are the smallest molecules with antigen-binding capacity and can therefore be expected to have properties different from conventional antibodies. In this study a library containing the VHH genes of a llama immunized with recombinant inner capsid protein VP6 was generated. Binders directed to VP6, in its native conformation within the viral particle, were selected and characterized. Four selected VHH directed to conformational epitopes of VP6 recognized all human and animal rotavirus strains tested and could be engineered for their use in immunodiagnostic tests for group A rotavirus detection. Three of the four VHH neutralized rotavirus in vivo independently of the strain serotype. Furthermore, this result was confirmed by in vivo partial protection against rotavirus challenge in a neonatal mouse model. The present study demonstrates for the first time a broad neutralization activity of VP6 specific VHH in vitro and in vivo. Neutralizing VHH directed to VP6 promise to become an essential tool for the prevention and treatment of rotavirus diarrhea.

Download Full-text