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Bibliographic Details
Main Authors:	Chen, Yiyu, Lian, Huiting, Liu, Guangming, Liu, Bin, Wei, Xiaofeng
Format:	Artículo científico
Language:	en
Published:	Analytical chemistry 2026
Subjects:	Biosensing Techniques Animals Allergens Immunoglobulin E Humans Biotinylation Epitope Mapping Circular Dichroism
Online Access:	https://pubmed.ncbi.nlm.nih.gov/42233754/
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Modular Avi-Tag Linkers Provide Predictable Protein Orientation for Generic Biosensor Construction. Chen, Yiyu Lian, Huiting Liu, Guangming Liu, Bin Wei, Xiaofeng Biosensing Techniques Animals Allergens Immunoglobulin E Humans Biotinylation Epitope Mapping Circular Dichroism Current multiplexed IgE assays suffer from random protein orientation, which reduces epitope accessibility. Here, we address this limitation by genetically fusing oyster allergens (Cra a 1 and Cra a 2) to an Avi-tag for site-specific biotinylation. The oriented capture probes, integrated with RCA-enhanced cell-free transcription, enable the simultaneous detection of two IgE targets with limits of detection of 1.5 and 27 pg/mL. Circular dichroism and epitope mapping reveal that Avi-tag conjugation maintains the native protein conformation, resulting in 2.3-fold higher IgE-binding activity compared with random NHS-biotinylation. Pilot testing with 20 serum samples shows 90% concordance with ELISA, demonstrating the potential for component-resolved allergy diagnosis.

if (filemtime($filename) + $maxlifetime < time()) { unlink($filename); $count++; } } return $count; } /** * A function that is called internally when session data is to be saved. * * @param string $sessId The current session ID * @param string $data The session data to write * * @return bool */ protected function saveSession($sessId, $data): bool { $sessFile = $this->path . '/sess_' . $sessId; if ($handle = fopen($sessFile, 'w')) { $return = false; // Lock the file for exclusive access to avoid issues with multiple // processes writing session simultaneously: if (flock($handle, LOCK_EX)) { $return = fwrite($handle, $data); // Make sure that there's no trailing data by truncating the file to // the correct length: ftruncate($handle, strlen($data)); fflush($handle); flock($handle, LOCK_UN); } fclose($handle); if ($return !== false) { return true; } } // If we got this far, something went wrong with the file output... // It is tempting to throw an exception here, but this code is called // outside of the context of exception handling, so all we can do is // echo a message.

// Anecdotal testing Today and Yesterday seems to indicate destroy() // is called by the garbage collector and everything is good. // Something to keep in mind though. return 0; } /** * Write function that is called when session data is to be saved. * * @param string $sessId The current session ID * @param string $data The session data to write * * @return bool */ public function write($sessId, $data): bool { if ($this->writesDisabled) { return true; } return $this->saveSession($sessId, $data); } /** * A function that is called internally when session data is to be saved. * * @param string $sessId The current session ID * @param string $data The session data to write * * @return bool */ abstract protected function saveSession($sessId, $data): bool; }

* * @return void */ public function writeClose() { // The assumption is that we're using PHP's ext/session. // session_write_close() will actually overwrite $_SESSION with an // empty array on completion -- which leads to a mismatch between what // is in the storage object and $_SESSION. To get around this, we // temporarily reset $_SESSION to an array, and then re-link it to // the storage object. // // Additionally, while you _can_ write to $_SESSION following a // session_write_close() operation, no changes made to it will be // flushed to the session handler. As such, we now mark the storage // object isImmutable. $storage = $this->getStorage(); if (! $storage->isImmutable()) { $_SESSION = $storage->toArray(true); session_write_close(); $storage->fromArray($_SESSION); $storage->markImmutable(); } } /** * Attempt to set the session name * * If the session has already been started, or if the name provided fails * validation, an exception will be raised. * * @param string $name * @return SessionManager * @throws Exception\InvalidArgumentException */ public function setName($name) { if ($this->sessionExists()) { throw new Exception\InvalidArgumentException( 'Cannot set session name after a session has already started'

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