<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>2304-0106</journal-id>
<journal-title><![CDATA[Anales de la Academia de Ciencias de Cuba]]></journal-title>
<abbrev-journal-title><![CDATA[Anales de la ACC]]></abbrev-journal-title>
<issn>2304-0106</issn>
<publisher>
<publisher-name><![CDATA[Academia de Ciencias de Cuba]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S2304-01062023000300020</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Mapeo epitópico fino y evolución dirigida del anticuerpo terapéutico nimotuzumab]]></article-title>
<article-title xml:lang="en"><![CDATA[Fine epitope mapping and directed evolution of the therapeutic antibody nimotuzumab]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Tundidor]]></surname>
<given-names><![CDATA[Yaima]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Rojas]]></surname>
<given-names><![CDATA[Gertrudis]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Pupo]]></surname>
<given-names><![CDATA[Amaury]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[García Hernández]]></surname>
<given-names><![CDATA[Claudia Patricia]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Centro de Inmunología Molecular  ]]></institution>
<addr-line><![CDATA[ La Habana]]></addr-line>
<country>Cuba</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2023</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2023</year>
</pub-date>
<volume>13</volume>
<numero>3</numero>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S2304-01062023000300020&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S2304-01062023000300020&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S2304-01062023000300020&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN  Introducción:  El nimotuzumab reconoce la región extracelular del receptor del factor de crecimiento epidérmico. A diferencia de otros anticuerpos en uso clínico, la interfaz de unión nimotuzumab-receptor se desconocía anteriormente. Este conocimiento permitiría comprender su mecanismo de acción y anticipar la posible resistencia al tratamiento por la pérdida del epitopo. La afinidad intermedia del nimotuzumab se ha invocado como explicación de su baja toxicidad, que permite que sea el único anticuerpo anti-receptor empleado en esquemas crónicos. A pesar de esta ventaja, su uso está limitado a tumores con alta expresión del receptor. La obtención de variantes del nimotuzumab con mayor afinidad e igual especificidad sería útil para entender la relación entre afinidad, toxicidad y efecto antitumoral. El objetivo fue caracterizar la interfaz funcional de unión entre el nimotuzumab y el receptor del factor de crecimiento epidérmico, y obtener variantes del nimotuzumab con mayor afinidad por el receptor que conservaran su especificidad epitópica fina.  Métodos:  Se desarrolló una estrategia de mapeo basada en la presentación sobre fagos filamentosos y mutagénesis extensiva del antígeno y de la región variable del nimotuzumab, validada por estudios de mutagénesis sobre proteínas producidas en células de mamífero. La maduración de la afinidad se realizó a partir de bibliotecas de fragmentos de anticuerpo presentados sobre fagos filamentosos.  Resultados:  Los residuos R353, S356, F357, T358 y H359 del dominio III del receptor, y la región hipervariable 3 de la cadena pesada del nimotuzumab fueron los principales contribuyentes a la unión. La evolución dirigida del paratopo culminó con 2 anticuerpos de mayor afinidad (3 veces y 3,6 veces), que conservaron la especificidad fina del nimotuzumab. Ambos tuvieron mayor actividad biológica que el anticuerpo parental. Las conclusiones fueron que el nimotuzumab reconoce un epitopo único respecto a otros anticuerpos aprobados en la clínica, y muestra una sensibilidad diferencial a las mutaciones del receptor. Las variantes del nimotuzumab de mayor afinidad brindan alternativas adicionales para el diagnóstico y tratamiento de tumores, además de herramientas útiles para estudiar la vinculación entre la afinidad y los efectos biológicos del anticuerpo.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT  Introduction:  Nimotuzumab recognizes the extracellular region of the epidermal growth factor receptor. Unlike other clinically approved antibodies, nimotuzumab/receptor binding interface was previously unknown. Such a knowledge would contribute to the understanding of its mechanisms of action and the prediction of treatment resistance due to epitope loss. It has been claimed that nimotuzumab intermediate affinity explains its lower toxicity, being the only anti-receptor antibody useful in chronic schemes. Despite this advantage, its use is limited to tumors displaying high receptor levels. Nimotuzumab variants with increased affinity and unchanged fine specificity would be useful to understand the link between affinity, toxicity and anti-tumor effects. The objective of this paper was to characterize the binding functional interface betwen the nimotuzumab and the receptor of the epidermic growth factor, and to obtain variants of nimotuzumab with higher affinity for the receptor that kept their fine epitope specificity.  Methods:  We developed a mapping strategy based on phage display and extensive mutagenesis of the target antigen and nimotuzumab variable region, validated by mutagenesis studies on proteins produced in mammalian cells. It was performed affinity maturation using libraries of phage-displayed antibody fragments.  Results:  Residues R353, S356, F357, T358 y H359 of receptor domain III and the third hypervariable region of nimotuzumab heavy chain were the main contributors to binding. Paratope evolution led to two antibodies with higher affinity (3 and 3.6 times) and the same fine specificity as compared to nimotuzumab. Both antibodies showed higher biological activity than the parental antibody. Conclusions, Nimotuzumab recognizes a unique epitope as compared to the other antibodies approved for clinical use, and shows differential sensitivity to receptor mutations. Higher affinity nimotuzumab variants offer additional alternatives for tumor diagnosis and treatment, as well as useful tools to explore the link between antibody affinity and biological effects.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[mapeo epitópico]]></kwd>
<kwd lng="es"><![CDATA[paratopo]]></kwd>
<kwd lng="es"><![CDATA[presentación sobre fagos]]></kwd>
<kwd lng="es"><![CDATA[mutagénesis]]></kwd>
<kwd lng="es"><![CDATA[maduración de la afinidad]]></kwd>
<kwd lng="en"><![CDATA[epitope mapping]]></kwd>
<kwd lng="en"><![CDATA[paratope]]></kwd>
<kwd lng="en"><![CDATA[phage display]]></kwd>
<kwd lng="en"><![CDATA[mutagenesis]]></kwd>
<kwd lng="en"><![CDATA[affinity maturation]]></kwd>
</kwd-group>
</article-meta>
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<year>2019</year>
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