No se puede utilizar, en forma directa, un GridView con una columna con TextBox con el atributo ReadOnly, para ello es necesario establecer este atributo en el evento RowDataBound del GridView, como sigue:
Private Sub GridView1_RowDataBound(ByVal sender As Object, ByVal e As System.Web.UI.WebControls.GridViewRowEventArgs) Handles GridView1.RowDataBound
Dim TxtBx As TextBox = e.Row.FindControl("TextBox1")
If TxtBx IsNot Nothing Then
TxtBx.Attributes.Add("readonly", "readonly")
End If
End Sub

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