Fluorosequencing, Essay Example

Fluorosequencing: Potential Applications in Drug Development and Biomarker Identification

Potential Applications or Advancement of Fluorosequencing

Applications

Parkinson’s disease is a condition that impacts about one million individuals in the U.S. (Eisyon, 2020). Due to the conditions such as aging and ineffective interventions, the disease has continued to challenge the medical industry. Fluorosequencing can be utilized to detect molecular biomarkers such as proteins. As a result, early intervention measures will be formed due to their high sensitivity to detecting a significant number of molecules from a small sample size. Early detection allows better intervention and treatments for the disease.

Fluorosequencing can be used for detecting cancerous cells. Antigens show a given cell’s health status (Eisyon, 2020). Cancerous cells or neoantigens are detected by methods such as T-cell receptor therapies. These therapies are based on the immunity’s capability to fight infections. Furthermore, these therapies lack the sensitivity for malignant cell detection since they require large sample sizes to be effective. Fluorosequencing could detect the peptides unique to cancerous cells in their early stages of development. It has high sensitivity detection, boosting the chances of treating cancer. The sensitivity occurs due to its capability to image single peptide molecules. A needle will be efficient in taking the small sample size from biopsy material of vital organs.

Additional areas of application of fluorosequencing include plant breeding, drug advancement, and industrial biology. Plants react to stressful conditions by modifying their protein compositions (Eisyon, 2020). Fluorosequencing can assess the change of plant proteins in different conditions to aid in plant breeding. Hence, plants can be modified to be resistant to adverse conditions. The impact of drugs on proteins is a vital study area in medicine. Fluorosequencing can reduce time consumption during tests to understand these reactions. Enzymes are used in various industrial processes, such as decontaminating consumables. Enzymes are safer than chemicals due to their non-toxicity. Fluoroseqenecing can be used to assess the behavior of enzymes in industrial processes. The process in which the peptide composition of enzymes shifts during industrial processes should be evaluated at different stages using fluorosequencing.

Advancing fluorosequencing

Fluorosequencing has shortcomings, such as relying on reagents that raise the degree of fluorescent dye decomposition. Some of these reagents consist of acids and pyridines. Due to this, the read time is minimized. Chemical labeling causes the peptide to experience partial sequencing. That leaves a remainder that must be compared with a base proteome (Alfaro et al., 2021). Inefficiencies of fluorosequencing can be caused by incorrect labeling of the base proteome. The labeling process of chemicals such as proteomes should be enhanced by using digitalized labeling to minimize errors.

Silica has been used to reduce the binding of proteins to glass for imaging procedures. Amine-derivatized surfaces are used with amide agents to stagnate proteins via carboxylic acids. Fluorosequencing of individual molecules of proteins uses the C-terminus or the surface binding agents, leading to issues with recognizing peptides. Protein identification can be improved by combining surface derivatizing agent 3-aminopropyl-triethoxysilane (APTES) and ordinary passivating components (Hinson et al., 2021). That will improve the coupling of proteins to the surfaces, aiding in protein identification.

Application of Fluorosequencing to Detecting Biomarkers of COVID-19 and Ocular Disease

COVID-19

A biomarker is an attribute assessed to depict the biological changes due to medical interventions. In pathology, peptides and proteins are used as biomarkers. Hence, these indicators can be used as biomarkers of COVID-19 (Jankowski, 2020). The biomarkers used to detect COVID-19 are linked with a rise in neutrophils, leukocytes, cytokine, tumor necrosis, and ferritin. However, biomarkers of serum ferritin, tropin I, and d-dimer have not been utilized due to sampling size challenges. Fluorosequencing can be fundamental in locating peptide or protein biomarkers of COVID-19 since it requires a small sample size.

Some viruses, such as COVID-19, are undetectable in asymptomatic individuals. The virus becomes unidentifiable when it undergoes the process of glycosylation (Eisyon, 2020). The virus uses sugars to cover its outline as camouflage. Due to this, the body is unable to respond to the virus. Fluorosequencing can be utilized to detect glycosylation trends of viruses as they are transmitted. The virus sample will have to be evaluated as they undergo glycosylation using fluorosequencing. Fluorosequencing utilizes single-molecule microscopy, fluorophore chemistry, and single-molecule microscopy techniques. The proteins are transformed into smaller peptides. They are placed on a glass surface through the terminus. A significant number of peptides labeled by fluorescents are examined in sequence. The shift in the fluorescent intensities causes the amino acids to be eliminated through Edman degradation (Alfaro et al., 2021). The final fluorescence readings allow any single peptides to be easily detected across a tremendous heterogeneity essential in glycosylation assessment. Afterwards, particular amino acids are labeled by easily identifiable fluorophores. As a result, of timely detection of COVID-19, proper responses can be formulated.

Ocular Disease

The eye is adaptive to several injuries or diseases due to structures such as cornea and sclera. Blood retinal barriers improve immunity while intraocular immune modulators reduce inflammation. Regardless of all these safety measures, the eyes are susceptible to various disorders. The disorders impact the eye matrix, leading to blindness (Tamhane et al., 2019). For instance, the anterior of the eye is susceptible to inflammation and infections. Meanwhile, the posterior of the eye is susceptible to conditions such as age-related macular degeneration and glaucoma. The treatment of these numerous conditions requires a comprehension of the disease and the physiological changes caused to the eyes. It is vital to use biomarkers to detect such conditions to advance drugs for treatment.

The feasible biomarkers for detecting ocular diseases should be extracted from the eyes. Ocular matrices that are feasible entail ocular tissue from cornea and tears. These matrices are suitable for detecting anterior eye complications. Tears have components such as proteins, electrolytes, and peptides (Tamhane et al., 2019). These components are good biomarkers since they are from numerous cells such as nerves and goblets. The tears’ biomarkers of proteins and inflammatory cytokines have been challenging to recognize by Edman and mass spectrometry techniques. That is due to the varying concentrations linked with different gathering and processing techniques. Due to this, comparing the different findings across various studies is difficult. Fluorosequencing will produce consistent results, using a few amino acids to recognize a protein sample. The change in proteome, lysosome and lactoferrin levels can be evaluated.

Furthermore, vitreous and aqueous humor should be used for establishing posterior defects. Accessing the posterior is challenging and requires clinical processes that may damage the eyes when too many samples are collected. Due to this, the small sample size requires an accurate detection method that will identify the biomarkers (Tamhane et al., 2019). Fluorosequencing will be suitable for assessing the small sample size because of its capability to assess fluorescence from single peptide molecules on a sample surface. The process can detect millions to billions of peptide molecules on a single slide, maximizing sample size.

References

Alfaro, J. A., Bohländer, P., Dai, M., Filius, M., Howard, C. J., van Kooten, X. F., … & Joo, C. (2021). The emerging landscape of single-molecule protein sequencing technologies. Nature methods, 18(6), 604-617.https://doi.org/10.1038/s41592-021-01143-1

Eisyon. (2020). Erisyon technologies are uniquely suited to address some key disease challenges. Eisyon. https://www.erisyon.com/applications/

Hinson, C. M., Bardo, A. M., Shannon, C. E., Rivera, S., Swaminathan, J., Marcotte, E. M., & Anslyn, E. V. (2021). Studies of Surface Preparation for the Fluorosequencing of Peptides. Langmuir, 37(51), 14856–14865. https://doi.org/10.1021/acs.langmuir.1c02644

Jankowski, J. (2020). Proteomic biomarkers to guide stratification for Covid?19 treatment: Exemplifying a path forward toward implementation?. Proteomics, 20(24), 2000229. https://doi.org/10.1002/pmic.202000229

Tamhane, M., Cabrera-Ghayouri, S., Abelian, G., & Viswanath, V. (2019). Review of biomarkers in ocular matrices: challenges and opportunities. Pharmaceutical Research, 36(3), 1-35. https://doi.org/10.1007/s11095-019-2569-8