Development of Recombinant Antibody by Yeast Surface Display Technology.

Recombinant antibodies have emerged as powerful tools in various fields, including therapeutics, diagnostics, and research applications. The selection of high-affinity antibodies with desired specificity is a crucial step in the development of recombinant antibody-based products. In recent years, yeast surface display technology has gained significant attention as a robust and versatile platform for antibody selection. This graphical review provides an overview of the yeast surface display technology and its applications in recombinant antibody selection. We discuss the key components involved in the construction of yeast surface display libraries, including the antibody gene libraries, yeast host strains, and display vectors. Furthermore, we highlight the strategies employed for affinity maturation and optimization of recombinant antibodies using yeast surface display. Finally, we discuss the advantages and limitations of this technology compared to other antibody selection methods. Overall, yeast surface display technology offers a powerful and efficient approach for the selection of recombinant antibodies, enabling the rapid generation of high-affinity antibodies for various applications.

Developing recombinant antibodies by phage display technology to neutralize viral infectious diseases.

The use of recombinant antibodies developed through phage display technology offers a promising approach for combating viral infectious diseases. By specifically targeting antigens on viral surfaces, these antibodies have the potential to reduce the severity of infections or even prevent them altogether. With the emergence of new and more virulent strains of viruses, it is crucial to develop innovative methods to counteract them. Phage display technology has proven successful in generating recombinant antibodies capable of targeting specific viral antigens, thereby providing a powerful tool to fight viral infections. In this mini-review article, we examine the development of these antibodies using phage display technology, and discuss the associated challenges and opportunities in developing novel treatments for viral infectious diseases. Furthermore, we provide an overview of phage display technology. As these methods continue to evolve and improve, novel and sophisticated tools based on phage display and peptide display systems are constantly emerging, offering exciting prospects for solving scientific, medical, and technological problems related to viral infectious diseases in the near future.

Nanobodies: A Game-Changer in Cell-Mediated Immunotherapy for Cancer.

Nanobodies are small, single-domain antibodies that have emerged as a promising tool in cancer immunotherapy. These molecules can target specific antigens on cancer cells and trigger an immune response against them. In this mini-review article, we highlight the potential of nanobodies in cell-mediated immunotherapy for cancer treatment. We discuss the advantages of nanobodies over conventional antibodies, their ability to penetrate solid tumors, and their potential to enhance the efficacy of other immunotherapeutic agents. We also provide an overview of recent preclinical and clinical studies that have demonstrated the effectiveness of nanobody-based immunotherapy in various types of cancer.

The reemergence of dengue virus in Sudan.

Dengue fever (DF) is a mosquito-transmitted arboviral disease caused by 1 of 4 closely related but antigenically distinct serotypes of dengue virus (DENV), DENV-1-4. The primary vector of DENV is Aedes aegypti and Aedes albopictus mosquitoes. Humans are the main carrier of the virus and the amplifying host with non-human primates plays a considerable role in sylvatic cycle. On November 8, 2022, an outbreak of dengue fever has killed at least five people in North Kordofan State. On 23 Nov 2022, the Sudanese Ministry of Health reported 3326 cases of dengue fever across 8 Sudanese States; while 23 patients died from the fever. Sudan is witnessing its worst outbreak of dengue fever in over a decade, especially in North and South Kordofan and Red Sea State are hit hard. In this review, we will focus on the recent outbreak of dengue fever in many Sudanese states.

Revolutionizing antiviral therapy with nanobodies: Generation and prospects.

As the world continues to grapple with infectious diseases, scientists are constantly searching for effective ways to combat these deadly pathogens. One promising avenue of research is the use of nanobodies as neutralization agents. These small proteins, derived from camelid antibodies, have several unique advantages over traditional antibodies, including their small size. Nanobodies are much smaller than conventional antibodies, typically weighing in at around 15 kDa compared to the 150 kDa of a typical human antibody. This small size allows them to penetrate into tight spaces that larger molecules cannot reach, such as the crevices on the surface of viruses or bacteria. This makes them highly effective at neutralizing viruses by binding to and blocking their key functional sites. In this mini-review we discuss the construction approaches of nanobodies, and some methods to increase the half-life of nanobodies. Moreover, we discuss Nanobodies and their therapeutic potential for infectious agents.

Novel multi epitope-based vaccine against monkeypox virus: vaccinomic approach.

While mankind is still dealing with the COVID-19 pandemic, a case of monkeypox virus (MPXV) has been reported to the WHO on May 7, 2022. Monkeypox is a viral zoonotic disease that has been a public health threat, particularly in Africa. However, it has recently expanded to other parts of the world, so it may soon become a global issue. Thus, the current work was planned and then designed a multi-epitope vaccine against MPXV utilizing the cell surface-binding protein as a target in order to develop a novel and safe vaccine that can evoke the desirable immunological response. The proposed MHC-I, MHC-II, and B-cell epitopes were selected to design multi-epitope vaccine constructs linked with suitable linkers in combination with different adjuvants to enhance the immune responses for the vaccine constructs. The proposed vaccine was composed of 275 amino acids and was shown to be antigenic in Vaxijen server (0.5311) and non-allergenic in AllerTop server. The 3D structure of the designed vaccine was predicted, refined and validated by various in silico tools to assess the stability of the vaccine. Moreover, the solubility of the vaccine construct was found greater than the average solubility provided by protein-Sol server which indicating the solubility of the vaccine construct. Additionally, the most promising epitopes bound to MHC I and MHC II alleles were found having good binding affinities with low energies ranging between - 7.0 and - 8.6 kcal/mol. According to the immunological simulation research, the vaccine was found to elicit a particular immune reaction against the monkeypox virus. Finally, the molecular dynamic study shows that the designed vaccine is stable with minimum RMSF against MHC I allele. We conclude from our research that the cell surface-binding protein is one of the primary proteins involved in MPXV pathogenesis. As a result, our study will aid in the development of appropriate therapeutics and prompt the development of future vaccines against MPXV.

SHERLOCK and DETECTR: CRISPR-Cas Systems as Potential Rapid Diagnostic Tools for Emerging Infectious Diseases.

Infectious diseases are one of the most intimidating threats to human race, responsible for an immense burden of disabilities and deaths. Rapid diagnosis and treatment of infectious diseases offers a better understanding of their pathogenesis. According to the World Health Organization, the ideal approach for detecting foreign pathogens should be rapid, specific, sensitive, instrument-free, and cost-effective. Nucleic acid pathogen detection methods, typically PCR, have numerous limitations, such as highly sophisticated equipment requirements, reagents, and trained personnel relying on well-established laboratories, besides being time-consuming. Thus, there is a crucial need to develop novel nucleic acid detection tools that are rapid, specific, sensitive, and cost-effective, particularly ones that can be used for versatile point-of-care diagnostic applications. Two new methods exploit unpredicted in vitro properties of CRISPR-Cas effectors, turning activated nucleases into basic amplifiers of a specific nucleic acid binding event. These effectors can be attached to a diversity of reporters and utilized in tandem with isothermal amplification approaches to create sensitive identification in multiple deployable field formats. Although still in their beginning, SHERLOCK and DETECTR technologies are potential methods for rapid detection and identification of infectious diseases, with ultrasensitive tests that do not require complicated processing. This review describes SHERLOCK and DETECTR technologies and assesses their properties, functions, and prospective to become the ultimate diagnostic tools for diagnosing infectious diseases and curbing disease outbreaks.

Immunoinformatics design of multiepitopes peptide-based universal cancer vaccine using matrix metalloproteinase-9 protein as a target.

A new approach toward cancer therapy is the use of cancer vaccine, yet the different molecular bases of cancers, reduce the effectiveness of this approach. In this article, we aim to use matrix metalloproteinase-9 protein (MMP9) which is an essential molecule in the survival and metastasis of all types of cancers as a target for universal cancer vaccine design. The reference sequence of MMP9 protein was obtained from NCBI databases. Furthermore, the B-cell and T cell-related peptides were analyzed using the IEDB website and other related soft wares. The best candidate peptides were then visualized using chimera software. Three peptides were found to be good candidates for interactions with B cells (SLPE, RLYT, and PALPR), while 10 peptides were found as good targets for interactions with MHC1 and another 10 peptides founded suitable for interactions with MHC2 with population coverages of 94.77 and 90.67%, respectively. Finally, the immune response simulation and molecular docking were done using the C-IMMSIM simulator and AutoDock Vina to confirm the effectiveness of the proposed vaccine. By the end of this project: twenty-three peptide-based vaccine was designed for use as a universal cancer vaccine which has a high world population coverage for MHC1 (94.77%) and MHC2 (90.67%) related alleles.

Cytokine Storm in COVID-19 Patients, Its Impact on Organs and Potential Treatment by QTY Code-Designed Detergent-Free Chemokine Receptors.

The novel coronavirus is not only causing respiratory problems, but it may also damage the heart, kidneys, liver, and other organs; in Wuhan, 14 to 30% of COVID-19 patients have lost their kidney function and now require either dialysis or kidney transplants. The novel coronavirus gains entry into humans by targeting the ACE2 receptor that found on lung cells, which destroy human lungs through cytokine storms, and this leads to hyperinflammation, forcing the immune cells to destroy healthy cells. This is why some COVID-19 patients need intensive care. The inflammatory chemicals released during COVID-19 infection cause the liver to produce proteins that defend the body from infections. However, these proteins can cause blood clotting, which can clog blood vessels in the heart and other organs; as a result, the organs are deprived of oxygen and nutrients which could ultimately lead to multiorgan failure and consequent progression to acute lung injury, acute respiratory distress syndrome, and often death. However, there are novel protein modification tools called the QTY code, which are similar in their structure to antibodies, which could provide a solution to excess cytokines. These synthetic proteins can be injected into the body to bind the excess cytokines created by the cytokine storm; this will eventually remove the excessive cytokines and inhibit the severe symptoms caused by the COVID-19 infection. In this review, we will focus on cytokine storm in COVID-19 patients, their impact on the body organs, and the potential treatment by QTY code-designed detergent-free chemokine receptors.

Epitope-Based Peptide Vaccine against Glycoprotein G of Nipah Henipavirus Using Immunoinformatics Approaches.

BACKGROUND: Nipah belongs to the genus Henipavirus and the Paramyxoviridae family. It is an endemic most commonly found at South Asia and has first emerged in Malaysia in 1998. Bats are found to be the main reservoir for this virus, causing disease in both humans and animals. The last outbreak has occurred in May 2018 in Kerala. It is characterized by high pathogenicity and fatality rates which varies from 40% to 70% depending on the severity of the disease and on the availability of adequate healthcare facilities. Currently, there are no antiviral drugs available for NiV disease and the treatment is just supportive. Clinical presentations for this virus range from asymptomatic infection to fatal encephalitis. OBJECTIVE: This study is aimed at predicting an effective epitope-based vaccine against glycoprotein G of Nipah henipavirus, using immunoinformatics approaches. METHODS AND MATERIALS: Glycoprotein G of the Nipah virus sequence was retrieved from NCBI. Different prediction tools were used to analyze the epitopes, namely, BepiPred-2.0: Sequential B Cell Epitope Predictor for B cell and T cell MHC classes II and I. Then, the proposed peptides were docked using Autodock 4.0 software program. Results and Conclusions. The two peptides TVYHCSAVY and FLIDRINWI have showed a very strong binding affinity to MHC class I and MHC class II alleles. Furthermore, considering the conservancy, the affinity, and the population coverage, the peptide FLIDRINWIT is highly suitable to be utilized to formulate a new vaccine against glycoprotein G of Nipah henipavirus. An in vivo study for the proposed peptides is also highly recommended.

Design of a Multiepitope-Based Peptide Vaccine against the E Protein of Human COVID-19: An Immunoinformatics Approach.

BACKGROUND: A new endemic disease has spread across Wuhan City, China, in December 2019. Within few weeks, the World Health Organization (WHO) announced a novel coronavirus designated as coronavirus disease 2019 (COVID-19). In late January 2020, WHO declared the outbreak of a "public-health emergency of international concern" due to the rapid and increasing spread of the disease worldwide. Currently, there is no vaccine or approved treatment for this emerging infection; thus, the objective of this study is to design a multiepitope peptide vaccine against COVID-19 using an immunoinformatics approach. METHOD: Several techniques facilitating the combination of the immunoinformatics approach and comparative genomic approach were used in order to determine the potential peptides for designing the T-cell epitope-based peptide vaccine using the envelope protein of 2019-nCoV as a target. RESULTS: Extensive mutations, insertion, and deletion were discovered with comparative sequencing in the COVID-19 strain. Additionally, ten peptides binding to MHC class I and MHC class II were found to be promising candidates for vaccine design with adequate world population coverage of 88.5% and 99.99%, respectively. CONCLUSION: The T-cell epitope-based peptide vaccine was designed for COVID-19 using the envelope protein as an immunogenic target. Nevertheless, the proposed vaccine rapidly needs to be validated clinically in order to ensure its safety and immunogenic profile to help stop this epidemic before it leads to devastating global outbreaks.

Extensive In Silico Analysis of ATL1 Gene : Discovered Five Mutations That May Cause Hereditary Spastic Paraplegia Type 3A.

BACKGROUND: Hereditary spastic paraplegia type 3A (SPG3A) is a neurodegenerative disease inherited type of Hereditary spastic paraplegia (HSP). It is the second most frequent type of HSP which is characterized by progressive bilateral and mostly symmetric spasticity and weakness of the legs. SPG3A gene mutations and the phenotype-genotype correlations have not yet been recognized. The aim of this work was to categorize the most damaging SNPs in ATL1 gene and to predict their impact on the functional and structural levels by several computational analysis tools. METHODS: The raw data of ATL1 gene were retrieved from dbSNP database and then run into numerous computational analysis tools. Additionally; we submitted the common six deleterious outcomes from the previous functional analysis tools to I-mutant 3.0 and MUPro, respectively, to investigate their effect on the structural level. The 3D structure of ATL1 was predicted by RaptorX and modeled using UCSF Chimera to compare the differences between the native and the mutant amino acids. RESULTS: Five nsSNPs out of 249 were classified as the most deleterious (rs746927118, rs979765709, rs119476049, rs864622269, and rs1242753115). CONCLUSIONS: In this study, the impact of nsSNPs in the ATL1 gene was investigated by various in silico tools that revealed five nsSNPs (V67F, T120I, R217Q, R495W, and G504E) are deleterious SNPs, which have a functional impact on ATL1 protein and, therefore, can be used as genomic biomarkers specifically before 4 years of age; also, it may play a key role in pharmacogenomics by evaluating drug response for this disabling disease.

In Silico Genetics Revealing 5 Mutations in CEBPA Gene Associated With Acute Myeloid Leukemia.

BACKGROUND: Acute myeloid leukemia (AML) is an extremely heterogeneous malignant disorder; AML has been reported as one of the main causes of death in children. The objective of this work was to classify the most deleterious mutation in CCAAT/enhancer-binding protein-alpha (CEBPA) and to predict their influence on the functional, structural, and expression levels by various Bioinformatics analysis tools. METHODS: The single nucleotide polymorphisms (SNPs) were claimed from the National Center for Biotechnology Information (NCBI) database and then submitted into various functional analysis tools, which were done to predict the influence of each SNP, followed by structural analysis of modeled protein followed by predicting the mutation effect on energy stability; the most damaging mutations were chosen for additional investigation by Mutation3D, Project hope, ConSurf, BioEdit, and UCSF Chimera tools. RESULTS: A total of 5 mutations out of 248 were likely to be responsible for the structural and functional variations in CEBPA protein, whereas in the 3'-untranslated region (3'-UTR) the result showed that among 350 SNPs in the 3'-UTR of CEBPA gene, about 11 SNPs were predicted. Among these 11 SNPs, 65 alleles disrupted a conserved miRNA site and 22 derived alleles created a new site of miRNA. CONCLUSIONS: In this study, the impact of functional mutations in the CEBPA gene was investigated through different bioinformatics analysis techniques, which determined that R339W, R288P, N292S, N292T, and D63N are pathogenic mutations that have a possible functional and structural influence, therefore, could be used as genetic biomarkers and may assist in genetic studies with a special consideration of the large heterogeneity of AML.

Novel Deleterious nsSNPs within MEFV Gene that Could Be Used as Diagnostic Markers to Predict Hereditary Familial Mediterranean Fever: Using Bioinformatics Analysis.

BACKGROUND: Familial Mediterranean Fever (FMF) is the most common autoinflammatory disease (AID) affecting mainly the ethnic groups originating from Mediterranean basin. We aimed to identify the pathogenic SNPs in MEFV by computational analysis software. METHODS: We carried out in silico prediction of structural effect of each SNP using different bioinformatics tools to predict substitution influence on protein structure and function. RESULT: 23 novel mutations out of 857 nsSNPs are found to have deleterious effect on the MEFV structure and function. CONCLUSION: This is the first in silico analysis of MEFV gene to prioritize SNPs for further genetic mapping studies. After using multiple bioinformatics tools to compare and rely on the results predicted, we found 23 novel mutations that may cause FMF disease and it could be used as diagnostic markers for Mediterranean basin populations.

Open repair of isolated traumatic subscapularis tendon tears with a synthetic soft tissue reinforcement.

BACKGROUND: Missed or chronic subscapularis tendon ruptures may have muscle atrophy and tendon retraction resulting in a large defect with high risk of re-tear after a surgical repair. To improve the clinical results of this challenging surgery, the repaired tendon could be augmented with endogenous or exogenous materials. The purpose of this study was to evaluate the structural tendon integrity and clinical outcomes after an open subscapularis tendon repair with a synthetic soft tissue reinforcement. MATERIALS AND METHODS: Ten patients were managed with an open repair of the subscapularis tendon with augmentation by means of SportMesh, a readily available synthetic degradable poly(urethaneurea) scaffold. Clinical findings were assessed for all patients preoperatively and postoperatively with use of the visual analog scale for pain and the DASH scoring system. All patients had an ultrasonographic study at the latest follow-up. RESULTS: The visual analog scale for pain (mean ± standard deviation) improved significantly (P < 0.01) from 7.9 ± 1.1 preoperatively to 1.95 ± 1.85 at the latest clinical follow-up evaluation. The mean DASH score at the latest clinical follow-up was 12.63 %. Ultrasound imaging revealed a structural intact repair at follow-up in 9 shoulders (90 %) with average 5.4 mm in thickness (4.3 mm in the contralateral healthy side). CONCLUSIONS: At a median follow-up of 23 months, 80 % (8 of 10) of patients had a good or excellent result after an open subscapularis tendon repair with a soft tissue reinforcement. As a synthetic material, SportMesh Soft Tissue Reinforcement eliminates the risk of collagen reactions, which may result from collagen or dermis patches. Although the follow-up is relatively short, our series shows a promising durable repair with a 10 % re-tear rate at an average of 23 postoperative months. Level of evidence Case Series, Treatment Study, Level IV.

Sonographic anatomy of the neck: The suprahyoid region.

The suprahyoid region extends from the base of the skull to the hyoid bone and includes the pharyngeal, parapharyngeal, parotid, carotid, masticator, retropharyngeal, and perivertebral spaces, as well as the oral cavity. The areas that can be explored by ultrasound include the parotid, carotid, and masticator spaces; the oral cavity; the submandibular and sublingual spaces; the floor of the mouth; and the root of the tongue. The parotid space contains the parotid gland and the excretory duct of Steno, the facial nerve, the external carotid artery, the retromandibular vein, and the intraparotid lymph nodes. The carotid space in the suprahyoid region of the neck contains the internal carotid artery, the internal jugular vein, cranial nerves IX to XII, and the sympathetic plexus. Only some parts of the masticator space can be explored sonographically: these include the masseter muscle, the zygomatic arch and the outer cortex of the ramus of the mandible, and the suprazygomatic portion of the temporalis muscle. The submandibular space houses the submandibular gland, the submental and submandibular lymph nodes, and the anterior belly of digastric muscle. The facial artery and vein and the lower loop of the hypoglossal nerve all pass through the submandibular space. The sublingual space includes the sublingual gland, the deep portion of the submandibular gland and its main excretory duct, the hypoglossal nerve (cranial nerve XII), the lingual nerve (branch of the mandibular branch of trigeminal), and the glossopharyngeal nerve (IX cranial nerve), and the lingual artery and vein. The mylohyoid muscle forms the floor of the mouth. The deepest portion of the oral tongue, the root, consists of the genioglossus and geniohyoid muscles and includes the septum of the tongue. In this article we present the ultrasound features of the structures located in the suprahyoid region of the neck.

Ultrasound anatomy of the neck: The infrahyoid region.

The infrahyoid region of the neck includes the visceral, anterior cervical, posterior cervical, carotid, retropharyngeal, and perivertebral spaces. The visceral space contains the thyroid, parathyroid glands, larynx, hypopharynx, the cervical trachea, and esophagus, the recurrent laryngeal nerve. The carotid space contains two parts, which extend from the skull base to the aortic arch and are delimited by the three layers of the deep cervical fascia (superficial, middle, and deep). It contains the internal carotid artery, the internal jugular vein, cranial nerves (IX-XII), the sympathetic plexus (suprahyoid compartment), the common carotid artery, the internal jugular vein, vagus nerve (infrahyoid compartment). The retropharyngeal space is a midlinespace containing adipose tissue that extends from the skull base to the upper mediastinum. It is located posterior to the pharynx and cervical esophagus, anterior to the danger area and the perivertebral space.THE PERIVERTEBRAL SPACE EXTENDS FROM THE SKULL BASE TO THE CLAVICLES AND INCLUDES TWO PARTS: prevertebral and paraspinal. The prevertebral space includes the prevertebral muscles (long muscles of the neck and head), the scalene muscles (anterior, middle, and posterior), the roots of the brachial plexus, the phrenic nerve, the vertebral arteries and veins, and the vertebral bodies. The paraspinal space contains the paraspinal muscles and the posterior elements of the cervical spine.The posterior cervical space is a deep space located posterior to the sternocleidomastoid muscle. It contains the accessory nerve, the accessory chain lymph nodes, the pre-axillary component of the brachial plexus, and the dorsal scapular nerve. The anterior cervical space is located deep to the strap muscles and sternocleidomastoid muscle. It surrounds the front and sides of the visceral space and is related posteriorly to the carotid space. This article reviews the ultrasound features of the structures located in the infrahyoid region of the neck.