Wenyu Li, Jun Chen, Isha Taneja, Wilson Edwards, Wenying Jian, Brian Geist
{"title":"Examining pre-analytical factors for bioanalysis of protein therapeutics crossing the blood-brain barrier through receptor-mediated transcytosis.","authors":"Wenyu Li, Jun Chen, Isha Taneja, Wilson Edwards, Wenying Jian, Brian Geist","doi":"10.1080/17576180.2026.2653856","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The blood-brain barrier (BBB) poses challenges for delivering large-molecule therapeutics to the brain. While engineered cross-BBB delivering platforms such as antibodies utilizing receptor-mediated transcytosis (RMT) aim to improve drug delivery, accurate quantization of antibody exposure in the brain is critical. This study investigates pre-analytical factors impacting the analysis of BBB-shuttling antibodies in mice and seeks to bring attention to steps that can be optimized to enhance efficiency and throughput.</p><p><strong>Research design and methods: </strong>We evaluated factors such as perfusion, capillary depletion, and freeze-thaw in a humanized mouse model treated with antibodies, with or without a BBB shuttle. Hemoglobin and albumin were investigated as markers for blood contamination.</p><p><strong>Results: </strong>Blood contamination was effectively reduced when cardiac puncture was performed, and perfusion further minimized the contamination. Albumin is a more representative blood contamination marker than hemoglobin for antibody bioanalysis in brain tissue. Capillary depletion did not significantly affect PK analysis under-tested conditions, and no considerable differences were found between frozen and fresh samples.</p><p><strong>Conclusions: </strong>Careful investigation of these pre-analytical procedures should be conducted to understand their impact. This will facilitate the development of an optimized and efficient workflow for discovery screening.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"245-253"},"PeriodicalIF":1.8000,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bioanalysis","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1080/17576180.2026.2653856","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2026/4/2 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
Background: The blood-brain barrier (BBB) poses challenges for delivering large-molecule therapeutics to the brain. While engineered cross-BBB delivering platforms such as antibodies utilizing receptor-mediated transcytosis (RMT) aim to improve drug delivery, accurate quantization of antibody exposure in the brain is critical. This study investigates pre-analytical factors impacting the analysis of BBB-shuttling antibodies in mice and seeks to bring attention to steps that can be optimized to enhance efficiency and throughput.
Research design and methods: We evaluated factors such as perfusion, capillary depletion, and freeze-thaw in a humanized mouse model treated with antibodies, with or without a BBB shuttle. Hemoglobin and albumin were investigated as markers for blood contamination.
Results: Blood contamination was effectively reduced when cardiac puncture was performed, and perfusion further minimized the contamination. Albumin is a more representative blood contamination marker than hemoglobin for antibody bioanalysis in brain tissue. Capillary depletion did not significantly affect PK analysis under-tested conditions, and no considerable differences were found between frozen and fresh samples.
Conclusions: Careful investigation of these pre-analytical procedures should be conducted to understand their impact. This will facilitate the development of an optimized and efficient workflow for discovery screening.
BioanalysisBIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL
CiteScore
3.30
自引率
16.70%
发文量
88
审稿时长
2 months
期刊介绍：
Reliable data obtained from selective, sensitive and reproducible analysis of xenobiotics and biotics in biological samples is a fundamental and crucial part of every successful drug development program. The same principles can also apply to many other areas of research such as forensic science, toxicology and sports doping testing.
The bioanalytical field incorporates sophisticated techniques linking sample preparation and advanced separations with MS and NMR detection systems, automation and robotics. Standards set by regulatory bodies regarding method development and validation increasingly define the boundaries between speed and quality.
Bioanalysis is a progressive discipline for which the future holds many exciting opportunities to further reduce sample volumes, analysis cost and environmental impact, as well as to improve sensitivity, specificity, accuracy, efficiency, assay throughput, data quality, data handling and processing.
The journal Bioanalysis focuses on the techniques and methods used for the detection or quantitative study of analytes in human or animal biological samples. Bioanalysis encourages the submission of articles describing forward-looking applications, including biosensors, microfluidics, miniaturized analytical devices, and new hyphenated and multi-dimensional techniques.
Bioanalysis delivers essential information in concise, at-a-glance article formats. Key advances in the field are reported and analyzed by international experts, providing an authoritative but accessible forum for the modern bioanalyst.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
